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Journal articles on the topic 'TRANSGENIC ANIMAL'

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Published: 4 June 2021

Last updated: 7 February 2022

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1

Oke, Krista B., Peter A. H. Westley, Darek T. R. Moreau, and Ian A. Fleming. "Hybridization between genetically modified Atlantic salmon and wild brown trout reveals novel ecological interactions." Proceedings of the Royal Society B: Biological Sciences 280, no. 1763 (July 22, 2013): 20131047. http://dx.doi.org/10.1098/rspb.2013.1047.

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Interspecific hybridization is a route for transgenes from genetically modified (GM) animals to invade wild populations, yet the ecological effects and potential risks that may emerge from such hybridization are unknown. Through experimental crosses, we demonstrate transmission of a growth hormone transgene via hybridization between a candidate for commercial aquaculture production, GM Atlantic salmon ( Salmo salar ) and closely related wild brown trout ( Salmo trutta ). Transgenic hybrids were viable and grew more rapidly than transgenic salmon and other non-transgenic crosses in hatchery-like conditions. In stream mesocosms designed to more closely emulate natural conditions, transgenic hybrids appeared to express competitive dominance and suppressed the growth of transgenic and non-transgenic (wild-type) salmon by 82 and 54 per cent, respectively. To the best of our knowledge, this is the first demonstration of environmental impacts of hybridization between a GM animal and a closely related species. These results provide empirical evidence of the first steps towards introgression of foreign transgenes into the genomes of new species and contribute to the growing evidence that transgenic animals have complex and context-specific interactions with wild populations. We suggest that interspecific hybridization be explicitly considered when assessing the environmental consequences should transgenic animals escape to nature.

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2

Santamaria, P. "Transgenic animal expressing diabetogenic Tcell receptor transgenes." Biofutur 1997, no. 167 (May 1997): 48. http://dx.doi.org/10.1016/s0294-3506(99)80369-x.

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3

Moore, Colin J., and T. Ben Mepham. "Transgenesis and Animal Welfare." Alternatives to Laboratory Animals 23, no. 3 (May 1995): 380–97. http://dx.doi.org/10.1177/026119299502300313.

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The two main techniques used in biomedical research for the production of transgenic animals have several implications for animal welfare in terms of the Three Rs of Russell & Burch. Some are intrinsic to the transgenic objectives, while others relate to the effects of mutations, transgene expression, associated methodologies, and husbandry or production systems. All of these actual and potential implications for animal welfare demand serious consideration within a broad ethical analysis of the technology. In the light, of the Three Rs, this may require a fundamental reappraisal of the processes by which such scientific procedures are approved.

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4

TSIKA, RICHARD W. "Transgenic Animal Models." Exercise and Sport Sciences Reviews 22, no. 1 (January 1994): 361–434. http://dx.doi.org/10.1249/00003677-199401000-00015.

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5

Towell, Jo. "Transgenic animal experiments." Nature Biotechnology 11, no. 9 (September 1993): 966. http://dx.doi.org/10.1038/nbt0993-966.

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6

Park, Frank. "Lentiviral vectors: are they the future of animal transgenesis?" Physiological Genomics 31, no. 2 (October 2007): 159–73. http://dx.doi.org/10.1152/physiolgenomics.00069.2007.

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Lentiviral vectors have become a promising new tool for the establishment of transgenic animals and the manipulation of the mammalian genome. While conventional microinjection-based methods for transgenesis have been successful in generating small and large transgenic animals, their relatively low transgenic efficiency has opened the door for alternative approaches, including lentiviral vectors. Lentiviral vectors are an appealing tool for transgenesis in part because of their ability to incorporate into genomic DNA with high efficiency, especially in cells that are not actively dividing. Lentiviral vector-mediated transgene expression can also be maintained for long periods of time. Recent studies have documented high efficiencies for lentiviral transgenesis, even in animal species and strains, such as NOD/ scid and C57Bl/6 mouse, that are very difficult to manipulate using the standard transgenic techniques. These advantages of the lentiviral vector system have broadened its use as a gene therapy vector to additional applications that include transgenesis and knockdown functional genetics. This review will address the components of the lentiviral vector system and recent successes in lentiviral transgenesis using both male- and female-derived pluripotent cells. The advantages and disadvantages of lentiviral transgenesis vs. other approaches to produce transgenic animals will be compared with regard to efficiency, the ability to promote persistent transgene expression, and the time necessary to generate a sufficient number of animals for phenotyping.

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7

Ju Kim, H., K. i. Naruse, W. S. Choi, K. S. Im, C. S. Park, and D. I. Jin. "332 ENHANCEMENT OF GROWTH PERFORMANCE IN DOUBLE TRANSGENIC MICE WITH GROWTH HORMONE RECEPTOR AND IGF-1 RECEPTOR GENES." Reproduction, Fertility and Development 17, no. 2 (2005): 317. http://dx.doi.org/10.1071/rdv17n2ab332.

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The effect of amplifying growth-related receptor signaling, through overexpression of receptors, on growth regulation in animals was examined. Transgenic mice lines were produced by DNA microinjection using the metallothionein promoter ligated to either the growth hormone receptor (GHR) or IGF-1 receptor (IGF-1R) genes (3 GHR founders and 3 IGF-1R founders). Transgenic mouse lines were estimated to contain approximately 4 to 20 copies of transgenes per cell by Southern blot analysis. Founder mice of each transgenic line transmitted transgenes into F1 and F2 pups with Mendelian ratio. Double transgenic (IGF-1R/GHR) mice were produced by the mating between nine pairs of IGF-1R and GHR hemizygous transgenic F1 mice. The transmission patterns in the 78 F2 pups produced from these matings were 20 with no transgene (25.6%), 17 with the IGF-1R gene (21.8%), 25 with the GHR gene (32.1%), and 16 with both GHR and IGF-1R genes (20.5%). The mRNA expression of transgenes using RT-PCR with the specific primers for IGF-IR and GHR genes was checked in tissues of transgenic mice. Double transgenic mice with IGF-IR and GHR genes expressed more mRNAs of transgenes than non-transgenic or single transgenic mice. Growth of double transgenic mice was fastest compared with single transgenic mice containing IGF-1R or GHR genes. And GHR transgenic mice grew faster than IGF-1R transgenic mice. When body weights of 15 transgenic mice for each transgenic line were measured at 4, 10, and 14 weeks after birth, double transgenic mice were significantly heavier compared with non-transgenic control mice at each stage (24 to 30% heavier in double transgenic mice; 15 to 20% heavier in single transgenic mice, P < 0.05). These results suggest that overexpression of growth-related receptor genes could promote the growth of transgenic animals with an additive effect.

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8

de Groot, Dorien M., Anton J. M. Coenen, Albert Verhofstad, François van Herp, and Gerard J. M. Martens. "In Vivo Induction of Glial Cell Proliferation and Axonal Outgrowth and Myelination by Brain-Derived Neurotrophic Factor." Molecular Endocrinology 20, no. 11 (November 1, 2006): 2987–98. http://dx.doi.org/10.1210/me.2006-0168.

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Abstract Brain-derived neurotrophic factor (BDNF) belongs to the neurotrophin family of neuronal cell survival and differentiation factors but is thought to be involved in neuronal cell proliferation and myelination as well. To explore the role of BDNF in vivo, we employed the intermediate pituitary melanotrope cells of the amphibian Xenopus laevis as a model system. These cells mediate background adaptation of the animal by producing high levels of the prohormone proopiomelanocortin (POMC) when the animal is black adapted. We used stable X. transgenesis in combination with the POMC gene promoter to generate transgenic frogs overexpressing BDNF specifically and physiologically inducible in the melanotrope cells. Intriguingly, an approximately 25-fold overexpression of BDNF resulted in hyperplastic glial cells and myelinated axons infiltrating the pituitary, whereby the transgenic melanotrope cells became located dispersed among the induced tissue. The infiltrating glial cells and axons originated from both peripheral and central nervous system sources. The formation of the phenotype started around tadpole stage 50 and was induced by placing white-adapted transgenics on a black background, i.e. after activation of transgene expression. The severity of the phenotype depended on the level of transgene expression, because the intermediate pituitaries from transgenic animals raised on a white background or from transgenics with only an approximately 5-fold BDNF overexpression were essentially not affected. In conclusion, we show in a physiological context that, besides its classical role as neuronal cell survival and differentiation factor, in vivo BDNF can also induce glial cell proliferation as well as axonal outgrowth and myelination.

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9

Robl, J. M., Z. Wang, P. Kasinathan, and Y. Kuroiwa. "Transgenic animal production and animal biotechnology." Theriogenology 67, no. 1 (January 2007): 127–33. http://dx.doi.org/10.1016/j.theriogenology.2006.09.034.

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10

Wigley, P., C. Becker, J. Beltrame, T. Blake, L. Crocker, S. Harrison, I. Lyons, et al. "Site-specific transgene insertion: an approach." Reproduction, Fertility and Development 6, no. 5 (1994): 585. http://dx.doi.org/10.1071/rd9940585.

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Methods to improve the production of transgenic animals are being developed. Conventional transgenesis, involving microinjection of DNA into fertilized eggs, has a number of limitations. These result from the inability to control both the site of transgene insertion and the number of gene copies inserted. The approach described seeks to overcome these problems and to allow single copy insertion of transgenes into a defined site in animal genomes. The method involves the use of embryonic stem cells, gene targeting and the FLP recombinase system.

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11

Wheeler, M. B., W. L. Hurley, S. J. Lane, G. E. Bressner, T. VanEtten, D. Kim, A. S. Lima, E. Monaco, and S. M. Wilson. "311 RISK ASSESSMENT OF α-LACTALBUMIN TRANSGENIC PIGS." Reproduction, Fertility and Development 20, no. 1 (2008): 235. http://dx.doi.org/10.1071/rdv20n1ab311.

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Assessment of the general risk posed by transgenic animals is important to their future contributions to society. Identification of potentially harmful properties of transgenic livestock is the initial step in a risk assessment. Direct and indirect impacts of potential harmful properties of transgenic livestock need to be evaluated at 3 levels: (1) characterization of how the transgene, the transgene product, and the transgenic livestock behave in their immediate environment; that is, in their barn or pen; (2) determination of possible impacts of large-scale release of transgenic livestock; that is, if they were to be integrated into the larger population of food animal livestock; and (3) determination of the more complex environmental and safety consequences of their release into the livestock population. We previously developed and characterized transgenic swine containing a mammary-specific transgene (bovine α-lactalbumin, bALAC) that results in increased milk production in sows (Bleck et al. 1998). We are currently determining whether bALAC is expressed in tissues of transgenic (T) swine other than the lactating mammary gland, and whether the transgene (DNA; Tg) crosses into non-transgenic control (C) swine under various physiological and physical conditions. The specific aims addressed in the present study were to determine (1) whether the Tg can be transferred directly from T animals to C animals by physical association or contact, and (2) whether the Tg can be transferred directly from an adult T animal to an adult C animal via mating. The T animals utilized in these studies were in generation 10 at least and have stable incorporation of the Tg. Comparable age- and weight-matched animals, T and C, were housed together allowing for general contact that is normal within swine production, for 180, 220, or 250 days after weaning. Due to the nature of swine behavior, these animals may ingest saliva, regurgitated food, and stool and urinary products as well as other bodily fluids and cells during normal housing and establishment of dominance hierarchy. In a second study, vaginal, cervical, uterine, oviductal, and ovarian tissues were collected from C females on 2 or 7 days after mating to T males. The presence of Tg in tissues from all C animals was tested via PCR. We have analyzed for the presence of the Tg in various tissues, including mammary gland, salivary gland, skin (sebaceous gland), muscle, lung, liver, kidney, brain, ovary, oviduct, uterus, cervix, vagina, and intestine. Preliminary results indicate no presence of the Tg in tissues of C animals (n = 20) after cohabitation for 180, 220, or 250 days (n = 201 samples analyzed) or at 2 (n = 3) or 7 (n = 5) days post-mating (n = 38 and 59 samples analyzed, respectively). This work provides a critical first step toward providing rigorous scientific data for risk assessment of transgenic livestock. The USDA BRAG Program, Project No. 2005–03799, supported this work.

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12

Choi, T., M. Huang, C. Gorman, and R. Jaenisch. "A generic intron increases gene expression in transgenic mice." Molecular and Cellular Biology 11, no. 6 (June 1991): 3070–74. http://dx.doi.org/10.1128/mcb.11.6.3070.

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To investigate the role of splicing in the regulation of gene expression, we have generated transgenic mice carrying the human histone H4 promoter linked to the bacterial gene for chloramphenicol acetyltransferase (CAT), with or without a heterologous intron in the transcription unit. We found that CAT activity is 5- to 300-fold higher when the transgene incorporates a hybrid intron than with an analogous transgene precisely deleted for the intervening sequences. This hybrid intron, consisting of an adenovirus splice donor and an immunoglobulin G splice acceptor, stimulated expression in a broad range of tissues in the animal. Although the presence of the hybrid intron increased the frequency of transgenics with significant CAT activity, it did not affect the integration site-dependent variation commonly seen in transgene expression. To determine whether the enhancement is a general outcome of splicing or is dependent on the particular intron, we also produced equivalent transgenics carrying the widely used simian virus 40 small-t intron. We found that the hybrid intron is significantly more effective in elevating transgene expression. Our results suggest that inclusion of the generic intron in cDNA constructs may be valuable in achieving high levels of expression in transgenic mice.

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13

Choi, T., M. Huang, C. Gorman, and R. Jaenisch. "A generic intron increases gene expression in transgenic mice." Molecular and Cellular Biology 11, no. 6 (June 1991): 3070–74. http://dx.doi.org/10.1128/mcb.11.6.3070-3074.1991.

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To investigate the role of splicing in the regulation of gene expression, we have generated transgenic mice carrying the human histone H4 promoter linked to the bacterial gene for chloramphenicol acetyltransferase (CAT), with or without a heterologous intron in the transcription unit. We found that CAT activity is 5- to 300-fold higher when the transgene incorporates a hybrid intron than with an analogous transgene precisely deleted for the intervening sequences. This hybrid intron, consisting of an adenovirus splice donor and an immunoglobulin G splice acceptor, stimulated expression in a broad range of tissues in the animal. Although the presence of the hybrid intron increased the frequency of transgenics with significant CAT activity, it did not affect the integration site-dependent variation commonly seen in transgene expression. To determine whether the enhancement is a general outcome of splicing or is dependent on the particular intron, we also produced equivalent transgenics carrying the widely used simian virus 40 small-t intron. We found that the hybrid intron is significantly more effective in elevating transgene expression. Our results suggest that inclusion of the generic intron in cDNA constructs may be valuable in achieving high levels of expression in transgenic mice.

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14

Smith, Charles. "Integration of transgenes in breeding programs." BSAP Occasional Publication 12 (1988): 71–80. http://dx.doi.org/10.1017/s0263967x00003293.

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ABSTRACTConventional animal breeding is in a healthy state, with good rates of genetic change possible, and economic efficiency being continually improved in the different livestock species. New reproductive techniques offer faster rates of change, as with the use of embryo transfer in sheep and cattle breeding. It is now possible to introduce foreign DNA, transgenes, into the germ line of farm animals and this offers new opportunities in the genetic improvement of economic merit. The transgenes should be directed at traits of high economic value, and be made in elite breeding stocks to benefit from past genetic improvements. Much still needs to be learned about engineering transgenes and about the basic biochemical-physiological processes they are designed to affect. However many transgenes will be produced over the next decade, and some may be useful in animal improvement. Founder transgene individuals are unique for incorporation site, copy number and expression, and will require screening, multiplication, testing and evaluation, in both hemizygous (TO) and homozygous (TT) form, for all economic traits. For a transgene with a large useful net effect on economic merit, the most rapid genetic response will come from fixing it in the elite breeding stock. An effect of 5-10 percent in economic merit will be needed to offset the normal genetic improvement displaced. Current experience with transgenes is of very large effects on the target trait, but with deleterious effects on fitness and economic merit. Control of the level, tissue and time of expression are crucial to the practical use of transgenes. With uncertainty in the results of a transgenic program, conventional programs will have to be maintained, so extra investment will be required. Patent rights may bring extra benefits to breeders, but there are also risks if the stock prove defective. Breeding strategies and organization may also change. Despite initial problems and uncertainties, the power of the transgenic methods is so great that the development of transgenic stocks is likely to become a very important tool in genetic improvement of livestock in the future.

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15

Powell, Ann, David Kerr, David Guthrie, and Robert Wall. "Lactation induction as a predictor of post-parturition transgene expression in bovine milk." Journal of Dairy Research 74, no. 2 (April 24, 2007): 247–54. http://dx.doi.org/10.1017/s0022029907002580.

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The bovine's long generation interval results in a delay of several years when evaluating mammary specific transgenes in genetically engineered animals. This experiment was conducted to evaluate the feasibility of reducing that waiting period. Lactation was induced in prepubertal bull and heifer calves as a means of predicting transgene behaviour during subsequent post-parturient lactations in the heifers themselves, and in daughters sired by the bulls. The animals carry a lactation-specific transgene encoding lysostaphin, an antimicrobial protein that kills Staphlococcus aureus, a mastitis-causing pathogen. Oestrogen, progesterone and dexamethasone were administered as previously described (Ball et al. 2000) to nine heifers (five transgenics) ranging in weight from 80 to 145 kg. Eight bull calves (seven transgenics) weighing 81–178 kg received additional oestrogen and progesterone injection prior to dexamethasone treatment. All nine heifers responded to the milk induction scheme yielding between 19 ml and 4·5 l over 5 d. Milk volume from the four responding males (30 μl to 2·5 ml) was significantly less than that harvested from females (P=0·025). Only bull calves >117 kg had a positive response. Lysostaphin was detected in all transgenic prepubertal heifers and in two transgenic prepubertal bull calves induced. A positive relationship was observed between lysostaphin's stapholytic activity in the two types of lactations (r2=0·907, P<0·001) thus providing a useful means of predicting subsequent lysostaphin production in post-partum milk.

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16

Wang, Yanli, Sihai Zhao, Liang Bai, Jianglin Fan, and Enqi Liu. "Expression Systems and Species Used for Transgenic Animal Bioreactors." BioMed Research International 2013 (2013): 1–9. http://dx.doi.org/10.1155/2013/580463.

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Transgenic animal bioreactors can produce therapeutic proteins with high value for pharmaceutical use. In this paper, we compared different systems capable of producing therapeutic proteins (bacteria, mammalian cells, transgenic plants, and transgenic animals) and found that transgenic animals were potentially ideal bioreactors for the synthesis of pharmaceutical protein complexes. Compared with other transgenic animal expression systems (egg white, blood, urine, seminal plasma, and silkworm cocoon), the mammary glands of transgenic animals have enormous potential. Compared with other mammalian species (pig, goat, sheep, and cow) that are currently being studied as bioreactors, rabbits offer many advantages: high fertility, easy generation of transgenic founders and offspring, insensitivity to prion diseases, relatively high milk production, and no transmission of severe diseases to humans. Noticeably, for a small- or medium-sized facility, the rabbit system is ideal to produce up to 50 kg of protein per year, considering both economical and hygienic aspects; rabbits are attractive candidates for the mammary-gland-specific expression of recombinant proteins. We also reviewed recombinant proteins that have been produced by targeted expression in the mammary glands of rabbits and discussed the limitations of transgenic animal bioreactors.

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17

Eyestone, WH. "Challenges and progress in the production of transgenic cattle." Reproduction, Fertility and Development 6, no. 5 (1994): 647. http://dx.doi.org/10.1071/rd9940647.

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The production of transgenic cattle presents a number of unique challenges not encountered in other species. First, the survival of microinjected zygotes is low; only 15% in vivo-derived develop into morulae and blastocysts and, of these, only about 18% yield live calves. Second, transgene integration frequency is relatively low, around 3%. Thus, more than 1000 zygotes must be injected to produce a single transgenic calf. Obtaining sufficient zygotes from donor cattle to sustain a transgenic cattle programme is logistically and financially prohibitive, since the average superovulated donor yields only about four microinjectable zygotes per collection attempt. In vitro oocyte maturation and fertilization techniques may be used to alleviate this problem, although initially the developmental potential of in vitro-derived microinjected zygotes is lower than their in vivo-produced counterparts (8% v. 15%, respectively, yield morulae and blastocysts). Since only 3-5% of calves born from microinjected zygotes produced in either fashion yield transgenics, at least 20-30 pregnancies must be carried to term for every transgenic calf born. These conditions require that large herds of donor and recipient cattle be maintained. Recipient requirements could be reduced if transgene integration frequency could be increased, but improvements in the near future are unlikely since the mechanism of integration after pronuclear microinjection is poorly understood. Alternatively, embryos could be screened for integrated transgenes before transfer; however, efforts in this area have been complicated by high frequencies of false positive results. Although yet to be developed, bovine embryonic stem cells would alleviate many of these problems and permit a wider range of genetic manipulations.

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18

Jura, Jacek, Zdzisław Smorąg, Barbara Gajda, Daniel Lipiński, and Ryszard Słomski. "Developmental Competence of CMV-Fut Transgenic and Non-Transgenic Pig Embryos Cultured in Vitro / Potencjał Rozwojowy CMV-Fut Transgenicznych I Nietransgenicznych Zarodków Świni Hodowanych In Vitro." Annals of Animal Science 13, no. 4 (September 1, 2013): 765–69. http://dx.doi.org/10.2478/aoas-2013-0051.

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Abstract Possible influence of a transgene on life functions of embryos makes it reasonable to confirm or deny it for a particular gene construct. In vitro development of an embryo is a widely used criterion of its competence. The aim of the study was to compare in vitro developmental capacity of transgenic and non-transgenic pig embryos. The results showed a statistically significant difference in in vitro developmental capacity of embryos obtained from transgenic and non-transgenic pigs. Developmental competence of embryos (morula and blastocyst stage) produced from zygotes obtained from transgenic sows decreased compared to that obtained from non-transgenic sows.

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19

Bishop, S. C., and J. A. Woolliams. "Utilization of the sex-determining region Y gene in beef cattle breeding schemes." Animal Science 53, no. 2 (October 1991): 157–64. http://dx.doi.org/10.1017/s0003356100020079.

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AbstractIn mammals ‘maleness’, i.e. the presence of testes, is thought to be controlled by a single gene on the Y chromosome. Recently, a candidate gene termed the SRY (sex-determining region Y) gene has been located. If the SRY gene is the gene causing maleness then a transgenic male with the SRY gene on an autosome would produce a greater proportion of male offspring than a normal male. This would be advantageous in situations where male offspring are more valuable than females. Such transgenic males have a reduced probability of propagating their genotype and an effort has to be made to avoid their extinction. This is at the cost of genetic progress which must be made to enable the transgenics to remain competitive with normal males.In a simulated beef cattle breeding scheme if half of the annual matings were made to transgenics then after 15 years of selection the transgenic males fell the equivalent of 2·6 years of selection behind males in a traditional herd. If all matings were made to transgenics they fell over 9 years behind. Selection for lean food conversion ratio was considered as an example. After 15 years of selection the gain in biological efficiency from more male offspring outweighed the loss from reduced genetic progress only when more than 0·5 of the bulls used in the breeding scheme were normal males. In practice, the difficulty of maintaining a small population of transgenic males along with other costs not included in the calculations suggest that breeding schemes in beef cattle with an SRY transgene would not be practicable without further technology.

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20

Zhen-Dan, Shi, Li Wan-Li, Zhang Yong-Liang, and Chen Xue-Jin. "Advances in the development of animal gene transfer." Chinese Journal of Agricultural Biotechnology 5, no. 2 (August 2008): 101–6. http://dx.doi.org/10.1017/s1479236208002179.

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AbstractEfficiency and specificity are key limiting factors for the production of transgenic animals. This review describes the recently developed animal gene transfer techniques, including non-site-specific methods of gene transfer into the testis and ovary for easy production of transgenic animals; gene targeting in embryonic stem cells, somatic cells and primordial germ cells for site-specific methods; methods to improve cloning efficiency in gene targeting; and site- and timing-specific gene targeting and controlled expression of transferred genes. In addition, methods of utilizing newly developed RNA interference, combined with the above techniques for controlling gene expression, to produce transgenic animals to spatio-temporally and reversibly knock down specific genes, are also discussed. The merits and disadvantages of each method are covered, as well as the potential use of these methods to develop transgenic animals for breeding new animal lines, to study disease models and to produce therapeutic medicines.

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21

Dixon, Bernard. "Transgenic Technology and Animal Welfare." Nature Biotechnology 13, no. 12 (December 1995): 1424. http://dx.doi.org/10.1038/nbt1295-1424.

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22

Götz, Jürgen. "Tau and transgenic animal models." Brain Research Reviews 35, no. 3 (July 2001): 266–86. http://dx.doi.org/10.1016/s0165-0173(01)00055-8.

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23

Boivin, Gregory P. "Update on transgenic animal technology." Lab Animal 44, no. 6 (May 19, 2015): 209. http://dx.doi.org/10.1038/laban.778.

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24

Rexroad, Caird E. "Transgenic technology in animal agriculture." Animal Biotechnology 3, no. 1 (January 1992): 1–13. http://dx.doi.org/10.1080/10495399209525759.

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25

Lee, Virginia M. Y., Theresa K. Kenyon, and John Q. Trojanowski. "Transgenic animal models of tauopathies." Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease 1739, no. 2-3 (January 2005): 251–59. http://dx.doi.org/10.1016/j.bbadis.2004.06.014.

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26

Kopp, J. B., and P. E. Klotman. "Transgenic animal models of renal development and pathogenesis." American Journal of Physiology-Renal Physiology 269, no. 5 (November 1, 1995): F601—F620. http://dx.doi.org/10.1152/ajprenal.1995.269.5.f601.

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The use of transgenic animals represents a powerful tool with which to address the role of particular gene products in vivo. Recent technical and biological advances have simplified the process of creating both transgenic mice and null-mutation mice. Increasing numbers of genetic control elements are available to direct transgene expression to particular renal cell types and to enhance the consistency of expression. These approaches have contributed significantly to our understanding of renal development and pathogenesis, in particular in the following areas: the roles of various oncogenes, homeobox genes, and growth factors in renal development and the pathogenesis of cystic renal diseases; the contribution of systemic and local expression of the renin-angiotensin system to blood pressure control; the role of growth factors and cytokines in progressive glomerular disease; the role of viral proteins in the pathogenesis of glomerular and tubular disease; and mechanisms of immune-mediated renal disease.

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Lee, S., H. Park, I. Kong, and Z. Wang. "30 A TRANSCRIPTION ACTIVATOR-LIKE EFFECTOR NUCLEASE (TALEN)-MEDIATED UNIVERSAL GENE KNOCK-IN STRATEGY FOR MAMMARY GLANDS-SPECIFIC EXPRESSION OF RECOMBINANT PROTEINS IN DAIRY CATTLE." Reproduction, Fertility and Development 26, no. 1 (2014): 129. http://dx.doi.org/10.1071/rdv26n1ab30.

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To harness the great capability of producing biologically active recombinant proteins with animal mammary glands, active research has been carried out in the past several decades to develop transgenic animals as bioreactors. However, when a transgene is introduced in the animal genome by random integration, the transgene tends to be subjected to epigenetic silencing, due to the so-called position effect from the chromatin environments surrounding the transgene integration sites, thereby resulting in low-level expression or total suppression. We report a universal transgenic strategy to knock in (KI) transgenes into the bovine β-casein gene locus allowing the expression of a transgene to be totally under the control of the endogenous regulatory sequences of the bovine β-casein gene. This universal KI strategy comprises two key components: one is the design of transcription activator-like effector nuclease (TALEN) constructs targeting the start codon region of bovine β-casein gene, and the other is the design of KI vectors in which a transgene of choice is flanked with homologous arms isolated from the ~500-bp bovine genomic DNAs immediately 5′ and 3′, respectively, of the translation start codon of the bovine β-casein gene. By using the human erythropoietin (hEPO) as the model transgene, we demonstrated that a transgene can be highly efficiently integrated immediately after the translation start codon of the bovine β-casein gene. In brief, the TALEN constructs were assembled by using the Golden Gate protocol. To KI the hEPO transgene, early passage (<5) of fibroblasts established from Holstein dairy cattle were cultured into full confluence in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS), harvested with 0.25% trypsin-EDTA, and co-transfected with KI vector and the TALEN constructs by the Amaxa Nucleofector system. For each experiment, 106 cells were transfected with 5 μg of KI vector and 5 μg of TALEN constructs. After 72 h post-transfection, cells were harvested and subjected to limiting dilution to obtain single-cell derived colonies. To screen for single-cell derived colonies carrying the correctly KI of hEPO in the β-casein locus, we performed genomic PCR amplifying the genomic junctions created by the KI of hEPO gene into the bovine genome. We identified and established 2 hEPO transgenic bovine fibroblast cell lines after screening 10 single-cell derived colonies from the transfected cells (20%). The genotype of these 2 colonies was also confirmed by sequencing the PCR products. We have initiated the effort to produce hEPO transgenic cattle by somatic cell nuclear transfer (SCNT), and the animal cloning results will be reported at the conference.

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Lipiński, Daniel, Joanna Zeyland, Andrzej Pławski, and Ryszard Słomski. "Determination of the Absolute Number of Transgene Copies in CMVFUT Transgenic Pigs." Annals of Animal Science 12, no. 3 (May 1, 2012): 349–56. http://dx.doi.org/10.2478/v10220-012-0029-z.

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Determination of the Absolute Number of Transgene Copies in CMVFUT Transgenic PigsThe aim of this research was to determine the number of transgene copies in the DNA of transgenic pigs. The copy number of the transgene was analysed in the transgenic animals with introduced pCMVFUT genetic construct containing a coding sequence of human H transferase under a control of CMV promoter. The copy number of the transgene that had integrated with the genome of the transgenic animals was analysed by qPCR with SYBR Green dye, which enabled nonspecific double-stranded DNA detection. CMVFT-2F and CMVFT-2R primers were used to amplify a 149 bp fragment of DNA. Forward primer had a sequence complementary to a promoter sequence and reverse primer to a coding sequence of H transferase. The copy number of the transgene in the examined samples was established by plotting the CT values obtained on a standard curve, which had been set by the usage of the CT values for the successive standard dilutions with known copy number (1.438-1.431 copies). As a standard we used pCMVFut genetic construct hydrolyzed with Not I restriction enzyme to a linear form. The real-time PCR results helped to establish the range of 3 - 4 as the number of the transgene copies that had integrated to the swine genome.

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Nowak-Imialek, M., W. A. Kues, B. Petersen, A. Lucas-Hahn, D. Herrmann, E. Lemme, M. Oropeza, J. W. Carnwath, and H. Niemann. "333 OCT-4 EXPRESSION ANALYSIS IN F0 AND F1 PORCINE OG2 TRANSGENICS." Reproduction, Fertility and Development 23, no. 1 (2011): 262. http://dx.doi.org/10.1071/rdv23n1ab333.

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OG2 transgenic pigs provide a new large animal model in which to study Oct-4 expression and the derivation, migration and maintenance of pluripotent cells. They may also prove to be a valuable tool for the development of cell-based therapies. The OG2 transgene consists of the genomic sequence of the murine Oct-4 gene with the enhanced green fluorescence protein (EGFP) reporter gene inserted between the promoter and the coding sequences. As previously reported, 11 OG2 founder animals were produced (7 male and 4 female). Two of the OG2-F0 transgenic boars were mated with 3 wild-type sows and with 2 OG2-F0 transgenic sows. The pregnancy of 1 wild-type sow was terminated at Day 5 after fertilization, and approximately 60% (14/23) of the flushed blastocysts expressed EGFP, demonstrating germ line transmission. The remaining 2 wild-type sows delivered 21 piglets, of which 11 were transgenic. The 2 OG2-F0 sows delivered 9 piglets, all of which were transgenic. Transgenesis and tissue-specific expression of the transgene were determined by Southern blotting, Northern blotting, and real-time PCR analysis. Germ cell-specific expression of the OG2 construct was confirmed in both F0 and F1 transgenics by fluorescence microscopy. Testis isolated from male transgenic piglets exhibited weak EGFP fluorescence in some cells within the seminiferous tubules, whereas testis tissue from adult transgenic boars gave strong EGFP expression in pre-spermatogonial cells. In contrast, fluorescence-activated cell sorting (FACS) analysis and fluorescence microscopy of ejaculated spermatozoa from 3 mature OG2-F0 boars displayed no EGFP fluorescence, as expected. Northern blot analysis of EGFP mRNA revealed stronger EGFP expression in the testis of adult transgenic pigs than in the testis from transgenic piglets. No EGFP mRNA was detected in other organs or in control testis isolated from wild-type piglets. Real-time PCR and Northern blot analysis showed that the time course and signal intensity of EGFP expression in OG2 testis paralleled expression of the endogenous Oct-4 gene in both transgenic and in wild-type testis, confirming that there is indeed stronger expression of Oct-4 in the adult testis than in testis from younger animals. We conclude that the OG2 founders exhibit germline transmission and that the offspring express EGFP in a pattern that faithfully mimics expression of the endogenous Oct-4 gene, thus providing a marker for pluripotent cells. We are currently using FACS to isolate EGFP-positive germ cells (pre-spermatogonial stem cells) from the testis of OG2 boars for further characterisation. Funded by BMBF.

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Kolb, Andreas. "Transgenic Animal Technology: Everything You Always Wanted to Know about Transgenics." Transgenic Research 13, no. 2 (April 2004): 201. http://dx.doi.org/10.1023/b:trag.0000026124.81947.03.

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Fischer, K. M. "Transgenic domestic animals provide an animal model for rheumatoid arthritis." Medical Hypotheses 38, no. 3 (July 1992): 240–43. http://dx.doi.org/10.1016/0306-9877(92)90102-i.

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BOHLENDER, JÜRGEN, DETLEV GANTEN, and FRIEDRICH C. LUFT. "Rats Transgenic for Human Renin and Human Angiotensinogen as a Model for Gestational Hypertension." Journal of the American Society of Nephrology 11, no. 11 (November 2000): 2056–61. http://dx.doi.org/10.1681/asn.v11112056.

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Abstract. Animal models of gestational hypertension are problematic. A novel mouse model was described earlier. The dams in that study were transgenic for human angiotensinogen and the sires for human renin; human renin was expressed in and produced by the placenta. This model was adapted to the rat, which has greater utility in terms of chronic instrumentation and physiologic measurements. Female rats transgenic for human angiotensinogen were mated with rats transgenic for human renin. Telemetry BP increased on day 5 of pregnancy from 110/80 mmHg to as high as 180/140 mmHg, while heart rate increased slightly. The renin transgene was expressed in the placenta, which resulted in increased human plasma renin concentration from 0 to 937 ± 800 ng angiotensin I ml/h; the values returned to 0 after delivery. Female rats transgenic for human renin that were mated with male rats transgenic for human angiotensinogen in contrast exhibited a decrease in BP. In these rats, human angiotensinogen in plasma remained undetectable. Double transgenic offspring of these transgenic rats developed hypertension and end-organ damage, regardless of the source of the transgenes. The conclusion is that transgenic rats that bear human renin and angiotensinogen genes make an attractive model for gestational hypertension. The rat model will have greater utility than the mouse model.

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Kong, Q. R., and Z. H. Liu. "427 INTEGRATION SITE ANALYSIS IN TRANSGENIC PIGS BY THERMAL ASYMMETRIC INTERLACED (TAIL)-PCR AND JUNCTION PCR." Reproduction, Fertility and Development 22, no. 1 (2010): 371. http://dx.doi.org/10.1071/rdv22n1ab427.

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Transgenic animals have been used to study gene function, produce important proteins, xenotransplantation donor, and generate models for the study of human diseases. Recent progress in animal cloning has provided an attractive alternative to improve transgenic efficiency, through the combination of transfection and somatic cell nuclear transfer (SCNT). However, when transgenic animals are produced by SCNT using randomly transfected cells as donor, the integration sites of transgene cannot be predicted. Many methods on the basis of genome walking have been demonstrated to clone transgene integration sites but they are either complicated or inefficient. In the study, we report a PCR-based method, thermal asymmetric interlaced PCR (TAIL-PCR), which relies on a series of 3 nested PCR reactions with transgene specific, designed with melting temperature of about 64, and arbitrary degenerate primers, by control of annealing temperature to efficiently reduce the nonspecific amplification to clone the integration sites in transgenic pigs by SCNT. Junction PCR combined with transgene-specific and integration site primers was performed to confirm the integration sites. Three integration sites were found (1 mapped on chromosome 4; the other 2 met a significant match in the pig expressed sequence tag database) in 2 founder transgenic pigs. Junction PCR resulted in specific amplification bands to identify the integration sites, and segregation of the integration sites was also detected in subsequent progeny by junction PCR analysis. We also used junction PCR combining with transgene-specific 5′ and 3′ integration site primers to analyze zygosity of the integration sites. Besides the specific amplification bands amplifying by transgene specific and integration site primers, bands amplified by 5′ and 3′ integration site primers were obtained to determine the heterozygosity of integration site. In conclusion, this strategy can be efficiently employed to clone transgene integration site and determine zygosity. This work was supported by grant from the State Transgenic Research Programme of China (Grant No. 2008ZX08006-002).

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Eaton, Heather. "Subjectivity and Suffering: Transgenic Animals, Christianity, and the need to Re-evaluate." Worldviews: Global Religions, Culture, and Ecology 14, no. 1 (2010): 26–56. http://dx.doi.org/10.1163/156853510x498041.

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AbstractThe many facets of transgenic animals are not addressed by secular or religious voices, and for many reasons; ignorance, absence of public debate, acceleration of the research, and apathy towards animals. There is a need to understand the basic parameters of transgenic animal research. Second, it is important to investigate Christian actual and potential responses, as well as grapple with the strengths and limits. Third, work in transgenic animals comes out of a deprived affective, aesthetic and ethical milieu where there is no rapport with animals as inherent subjects. With new insights from religion and animal studies, it may be possible to transform the prevailing utilitarian view of animals.

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Ahn, K. S., M. Kwon, B. C. Koo, J. Y. Won, S. Y. Heo, T. Kim, and H. Shim. "20 PRODUCTION OF NUCLEAR TRANSFER EMBRYOS FROM PORCINE FETAL FIBROBLAST CELLS CARRYING A TETRACYCLINE-INDUCIBLE TRANSGENE." Reproduction, Fertility and Development 18, no. 2 (2006): 118. http://dx.doi.org/10.1071/rdv18n2ab20.

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Constitutive expression of A transgene often results in serious physiological disturbances in transgenic animals. For instance, systemic overexpression of human growth hormone in transgenic pigs has resulted in detrimental side effects in general health and reproductive performance. One of the solutions to such problem would be inducible expression of a transgene that may restrict production of foreign proteins from transgenic animals only when needed. In this study, a retrovirus vector was designed to express the green fluorescent protein (GFP) gene under the control of the tetracycline-inducible promoter. Transformation of porcine fetal fibroblast cells was achieved by infection of the cells with the vector and subsequent antibiotic selection. To induce transgene expression, transformed porcine fetal fibroblast cells were cultured in medium supplemented with doxycycline for 48 h. Induction of the GFP gene was verified by the emission of fluorescence from transformed cells. Nuclei of transformed cells with or without doxycycline treatment were transferred into enucleated oocytes, and the induction efficiency was analyzed by monitoring fluorescent emission during development of reconstituted embryos to the blastocyst stage. In addition, differences in the rates of blastocyst development between experimental groups were analyzed by Student's t-test. Blastocyst formation of nuclear transfer embryos using transformed cells with tetracycline-inducible retrovirus vector (12.0%, 128/1072) was not significantly different (P > 0.05) from that with non-inducible control vectors (13.7%, 41/300), suggesting that an introduction of tetracycline-inducible retrovirus vector was not particularly harmful to the development of nuclear transfer embryos. Also, the blastocyst development rate of nuclear transfer embryos after induction of transgene by doxycycline (12.1%, 99/815) was not significantly different (P > 0.05) from that of the non-induced counterparts (11.3%, 29/257), suggesting that the induction of transgene did not affect the development of transgenic clone embryos. In a majority of embryos, high expression of the GFP gene was observed in cloned embryos with transgene induction, whereas poor or no GFP expression was detected in non-induced controls. The results from this study suggest that tetracycline-inducible expression of transgenes in nuclear transfer embryos may be used for production of foreign proteins in transgenic animals in a more controlled manner than with conventional procedures. Further experiments on transfer of cloned embryos carrying such an inducible transgene to recipients may enable production of transgenic pigs with fewer side effects from unregulated expression of the transgene.

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Urano, K., N. Tamaoki, and T. Nomura. "Establishing a Laboratory Animal Model From a Transgenic Animal." Veterinary Pathology 49, no. 1 (December 6, 2011): 16–23. http://dx.doi.org/10.1177/0300985811430318.

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37

Gama, L. T., C. Smith, and J. P. Gibson. "Transgene effects, introgression strategies and testing schemes in pigs." Animal Science 54, no. 3 (June 1992): 427–40. http://dx.doi.org/10.1017/s0003356100020894.

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AbstractStrategies of introgressing a transgene into a pig nucleus undergoing mass selection for net merit were deterministically evaluated. They consisted of systematically backcrossing hemizygous transgenic sires to females from the selection nucleus, or vice versa, followed by intercrossing of hemizygous individuals, assuming different levels of heritability (h2), polygenic breeding value of the founder animal, inbreeding depression and constraints in availability of resources. The polygenic breeding value of the founder transgenic animal and inbreeding depression were of negligible importance if backcrossing lasted for at least three generations, and there was little advantage in extending backcrossing much further. The best introgression strategy examined was to backcross selection nucleus sires to hemizygous females, but this was a less efficient strategy in terms of testing transgene effects. Testing the survival of homozygous carriers requires approximately five and 100 matings among hemizygous individuals to detect a reduction in viability of 0·5 and 0·1, respectively. Comparing several candidate transgenes in the first generations of backcrossing is feasible, and does not result in substantial delays in improvement of polygenic breeding value in the selected transgene. If resources are limited, the magnitude of the transgene effect (as a proportion of the mean) that compensates for the genetic lag incurred by its introgression is about 0·1 for most economic traits in pigs. To compensate for less selection while backcrossing and for risk in use, transgenes must have an appreciable effect on economic merit to make their introgression worthwhile, even when the additional costs of transgene production are ignored.

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38

Powell, A., D. Kerr, and R. Wall. "379 LACTATION INDUCTION IN PREPUBERTAL BULLS AND HEIFERS AS A TOOL FOR PREDICTING MAMMARY SPECIFIC TRANSGENE EXPRESSION IN CATTLE." Reproduction, Fertility and Development 18, no. 2 (2006): 296. http://dx.doi.org/10.1071/rdv18n2ab379.

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The bovine's long generation interval results in a lapse, from the time of birth, of two to three years before mammary-specific transgenes can be assessed in genetically engineered animals. This experiment was conducted in an attempt to reduce that waiting period by up to two years. Lactation was induced in prepubertal bull and heifer calves, 3 to 8 mo of age, as a means of predicting transgene behavior during subsequent normal lactations in the heifers and daughters of bulls. Transgenic animals tested were either founder animals, produced by somatic cell nuclear transfer, or G1 offspring of founder bulls. The transgene consists of a lactation specific sequence encoding lysostaphin, an antimicrobial protein targeted against Staphylococcus aureus, a mastitis-causing pathogen. Estrogen, progesterone, and dexamethasone were administered as previously described (Ball et al. 2000 J. Dairy Sci. 83, 2459) to nine heifers (transgenics = 5) ranging in weight from 90 to 165 kg. Eight bull calves (transgenics = 7) weighing from 81 to 178 kg received additional estrogen and progesterone injections as well as reserpine prior to dexamethasone treatment. Animals were hand-milked twice daily for 4 to 7 days. All nine heifers responded to the milk induction scheme, yielding between 19 mL and 4.5 L. Milk volume from the three responding males (100 �L to 2.5 mL) was significantly less than that harvested from females (P = 0.025). Only bull calves over 150 kg had a positive response. Transgenic females produce less milk then non-transgenics (313 � 494 vs. 2276 � 552 mL, respectively; P = 0.033). Most importantly, there was no detectable difference between the concentration of lysostaphin in milk from induction (8.1 � 2.7 �g/mL) and natural lactations (3.5 � 2.6 �g/mL) in the four transgenic heifers tested (P = 0.229). The result was the same when lysostaphin was analyzed as a percentage of total protein (P = 0.427). Induction of a G1 heifer and a bull calf from the same founder bull produced similar lysostaphin concentrations in their milk (5.6 � 0.9 and 5.2 � 0.5 �g/mL; respectively). �-lactoglobulin concentration was also similar during induced and natural lactation (P = 0.165) for all animals studied. However, total protein was greater in induced milk samples compared to natural lactation samples (28.4 � 1.7 vs. 21.2 � 1.7 mg/mL; P = 0.007) as was lactoferrin (707 � 51 vs. 213 � 51 �g/mL; P < 0.001). Conversely, compared to that in induced milk samples, lactose was more concentrated in the natural lactation samples (34.6 � 2.5 vs. 46.0 � 2.1 g/L). In this study transgene expression was detected in milk from induced lactations and its concentration in those samples was generally predictive of product concentration in the natural lactation milk. The induction protocol was effective in male (>150 kg) and female calves.

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SUN, Zhen-Hong, Xiang-Yang MIAO, and Rui-Liang ZHU. "New advances in animal transgenic technology." Hereditas (Beijing) 32, no. 6 (July 22, 2010): 539–47. http://dx.doi.org/10.3724/sp.j.1005.2010.00539.

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40

Paul, Martin, and Wolfgang-Michael Franz. "Transgenic animal models for hypertension research." Trends in Cardiovascular Medicine 5, no. 3 (May 1995): 108–14. http://dx.doi.org/10.1016/1050-1738(95)00053-c.

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41

Luboń, Henryk, Rekha K. Paleyanda, William H. Velander, and William N. Drohan. "Blood proteins from transgenic animal bioreactors." Transfusion Medicine Reviews 10, no. 2 (April 1996): 131–43. http://dx.doi.org/10.1016/s0887-7963(96)80089-7.

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42

Aldhous, Peter. "Europe approves first transgenic animal patent." Nature 353, no. 6345 (October 1991): 589. http://dx.doi.org/10.1038/353589a0.

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43

FUKUCHI, KEN-ICHIRO, CHARLES E. OGBURN, ANNETTE C. SMITH, DENNIS D. KUNKEL, CLEMENT E. FURLONG, SAMIR S. DEEB, DAVID NOCHLIN, S. MARK SUMI, and GEORGE M. MARTIN. "Transgenic Animal Models for Alzheimer's Diseasea." Annals of the New York Academy of Sciences 695, no. 1 (September 1993): 217–23. http://dx.doi.org/10.1111/j.1749-6632.1993.tb23055.x.

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Li, Xiaojiang, and Shihua Li. "Neuropathology of transgenic HD animal models." Molecular Neurodegeneration 7, Suppl 1 (2012): L17. http://dx.doi.org/10.1186/1750-1326-7-s1-l17.

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45

Gordon, J. W. "Transgenic Technology and Laboratory Animal Science." ILAR Journal 38, no. 1 (January 1, 1997): 32–41. http://dx.doi.org/10.1093/ilar.38.1.32.

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46

Fox, Jeffrey L. "FDA transgenic animal guidance finally surfaces." Nature Biotechnology 26, no. 11 (November 2008): 1205–6. http://dx.doi.org/10.1038/nbt1108-1205.

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47

Barrett, Graham, and John J. Mullins. "Transgenic animal models of cardiovascular disease." Current Opinion in Biotechnology 3, no. 6 (December 1992): 637–40. http://dx.doi.org/10.1016/0958-1669(92)90008-7.

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48

Jeong, Y. H., G. H. Jang, I. S. Hwang, C. H. Park, H. J. Lee, Y. W. Jeong, S. H. Hyun, et al. "330 REDUCED HYPERACUTE REJECTION BY TRIPLE TRANSGENIC EXPRESSION OF HUMAN COMPLEMENT REGULATORY FACTORS (hDAF and hCD59) AND H-TRANSFERASE." Reproduction, Fertility and Development 23, no. 1 (2011): 261. http://dx.doi.org/10.1071/rdv23n1ab330.

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The present study was conducted to establish a porcine transgenic cell line with human CRPs and HT genes, focused on hyperacute rejection (HAR) considering clinical xenotransplantation as alternative sources of human organs. As a first step towards establishing the stable cell line, the cDNA for 3 genes encoding human DAF, CD59, and H-transferase were cloned and sequenced. A tricistronic expression vector was constructed with the aid of 2 IRES elements (pCMV-hDAF_IRES-hHT_IRES-hCD59). The CMV-based expression vector was then introduced into miniature pig ear fibroblast cells by electroporation. Reverse transcription PCR analysis revealed that cell lines stably expressing human transgene-specific transcripts were established. The inhibitory effect of immune response in the established transgenic cell lines was measured by human serum-mediated cytolysis assay, as measured by ELISA. Under the assay conditions (based on human serum from 10 to 50%), the transgenic cell group showed significantly greater survival rate under various serum concentrations than did the nontransgenic cell control group. Moreover, the transgenic cell lines used as nuclear donors for a subsequent NT experiment were confirmed to be expressing their transgene transcripts in vitro developed preimplantation stage embryos. These results indicated that the established cell lines with human transgenes might have an inhibitory effect against lysis by human complement. It is possible that these transgenic cells could serve as nuclear donors to produce transgenic cloned pigs for xenotransplantation.

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Houdebine, L.-M. "Use of Transgenic Animals to Improve Human Health and Animal Production." Reproduction in Domestic Animals 40, no. 4 (August 2005): 269–81. http://dx.doi.org/10.1111/j.1439-0531.2005.00596.x.

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50

Nyqvist, Daniel, Göran Mattsson, Martin Köhler, Varda Lev-Ram, Arne Andersson, Per-Ola Carlsson, Astrid Nordin, Per-Olof Berggren, and Leif Jansson. "Pancreatic islet function in a transgenic mouse expressing fluorescent protein." Journal of Endocrinology 186, no. 2 (August 2005): 333–41. http://dx.doi.org/10.1677/joe.1.06204.

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Pancreatic islet function and glucose homeostasis have been characterized in the transgenic YC-3.0 mouse, which expresses the yellow chameleon 3.0 (YC-3.0) protein under the control of the β-actin and the cytomegalovirus promoters. Fluorescence from the enhanced yellow fluorescent protein (EYFP), one part of the yellow chameleon protein, was used as a reporter of transgene expression. EYFP was expressed in different quantities throughout most cell types, including islet endocrine and stromal cells. No adverse effects of the transgene on animal health, growth or fertility were observed. Likewise, in vivo glucose homeostasis, mean arterial blood pressure and regional blood flow values were normal. Furthermore, the transgenic YC-3.0 mouse had a normal β-cell volume and mass as well as glucose-stimulated insulin release in vitro, compared with the C57BL/6 control mouse. Isolated islets from YC-3.0 animals continuously expressed the transgene and reversed hyperglycemia when transplanted under the renal capsule of alloxan-diabetic nude mice. We conclude that isolated pancreatic islets from YC-3.0 animals implanted into recipients without any EYFP expression, constitute a novel and versatile model for studies of islet engraftment.

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