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Journal articles on the topic 'Replikon'

Author: Grafiati

Published: 4 June 2025

Last updated: 1 August 2025

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1

Şahiner, Fatih, and İsmail Selçuk Aygar. "A New Era in Vaccine Technology: mRNA-Based Vaccine Design." Journal of Molecular Virology and Immunology 1, no. 3 (2021): 9–17. https://doi.org/10.46683/jmvi.2020.15.

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<strong>Özet</strong> Messenger RNA (mRNA) teknolojisi hem genetik hastalıkların ve kanserlerin tedavisinde (terapötik kanser aşıları) hem de enfeksiyöz hastalıkların yayılımının önlenmesinde gelecek vaat eden yeni nesil bir yaklaşımı temsil eder. mRNA aşı sistemlerinin temel mantığı istenilen bir proteinin viral bir enfeksiyonu taklit ederek vücutta üretilmesini sağlamak ve onun işlevlerinden yararlanmaktır. DNA temelli sistemlerden ve viral vektörlerden farklı olarak üretilmek istenilen proteine ait genetik kodu taşıyan mRNA molekülleri ikinci bir

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2

Alangari, Abdulaziz, Ahmad Abu Jaffal, Naif Almutairi, and Abdullah A. Alyousef. "Plasmid Replicon Diversity of Clinical Uropathogenic Escherichia coli Isolates from Riyadh, Saudi Arabia." Journal of Pure and Applied Microbiology 16, no. 1 (2022): 540–48. http://dx.doi.org/10.22207/jpam.16.1.51.

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The aim of this study was to identify and compare the plasmid replicons of clinical uropathogenic Escherichia coli (UPEC) isolates, involving extended spectrum β-lactamase (ESBL)-positive and ESBL-negative, E. coli ST131 and non-ST131 and various ST131 subclones. Plasmid replicon typing on 24 clinical UPEC isolates was carried out using polymerase chain reaction-based replicon typing. A statistical analysis was performed to assess the associations between plasmid replicon types and ESBL carriage, and to evaluate the link between ST131 isolates and high replicon carriage. Eight replicons, I1α,

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3

Kato, Nobuyuki, Takashi Nakamura, Hiromichi Dansako, et al. "Genetic variation and dynamics of hepatitis C virus replicons in long-term cell culture." Journal of General Virology 86, no. 3 (2005): 645–56. http://dx.doi.org/10.1099/vir.0.80479-0.

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Hepatitis C virus (HCV) genomic sequences are known to vary widely among HCV strains, but to date there have been few reports on the genetic variations and dynamics of HCV in an experimental system of HCV replication. In this study, a genetic analysis of HCV replicons obtained in long-term culture of two HCV replicon cells (50-1 and 1B-2R1), which were established from two HCV strains, 1B-1 and 1B-2, respectively, was performed. One person cultured 50-1 cells for 18 months, and two people independently cultured 50-1 cells for 12 months. 1B-2R1 cells were also cultured for 12 months. The whole

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4

Wu, Joh-Sin, Ju-Ying Kan, Hsueh-Chou Lai, and Cheng-Wen Lin. "Development of Zika Virus Mini-Replicon Based Single-Round Infectious Particles as Gene Delivery Vehicles." Viruses 15, no. 8 (2023): 1762. http://dx.doi.org/10.3390/v15081762.

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Zika virus (ZIKV) is a type of RNA virus that belongs to the Flaviviridae family. We have reported the construction of a DNA-launched replicon of the Asian-lineage Natal RGN strain and the production of single-round infectious particles (SRIPs) via the combination of prM/E virus-like particles with the replicon. The main objective of the study was to engineer the ZIKV replicon as mammalian expression vectors and evaluate the potential of ZIKV mini-replicon-based SRIPs as delivery vehicles for heterologous gene expression in vitro and in vivo. The mini-replicons contained various genetic elemen

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5

Murray, Edward M., Jay A. Grobler, Eric J. Markel та ін. "Persistent Replication of Hepatitis C Virus Replicons Expressing the β-Lactamase Reporter in Subpopulations of Highly Permissive Huh7 Cells". Journal of Virology 77, № 5 (2003): 2928–35. http://dx.doi.org/10.1128/jvi.77.5.2928-2935.2003.

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ABSTRACT Progress toward development of better therapies for the treatment of hepatitis C virus (HCV) infection has been hampered by poor understanding of HCV biology and the lack of biological assays suitable for drug screening. Here we describe a powerful HCV replication system that employs HCV replicons expressing the β-lactamase reporter (bla replicons) and subpopulations of Huh7 cells that are more permissive (or “enhanced”) to HCV replication than naïve Huh7 cells. Enhanced cells represent a small fraction of permissive cells present among naïve Huh7 cells that is enriched during selec

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6

Jackson, Dean A., and Ana Pombo. "Replicon Clusters Are Stable Units of Chromosome Structure: Evidence That Nuclear Organization Contributes to the Efficient Activation and Propagation of S Phase in Human Cells." Journal of Cell Biology 140, no. 6 (1998): 1285–95. http://dx.doi.org/10.1083/jcb.140.6.1285.

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In proliferating cells, DNA synthesis must be performed with extreme precision. We show that groups of replicons, labeled together as replicon clusters, form stable units of chromosome structure. HeLa cells were labeled with 5-bromodeoxyuridine (BrdU) at different times of S phase. At the onset of S phase, clusters of replicons were activated in each of ∼750 replication sites. The majority of these replication “foci” were shown to be individual replicon clusters that remained together, as stable cohorts, throughout the following 15 cell cycles. In individual cells, the same replication foci we

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7

Graziani, Rita, and Giacomo Paonessa. "Dominant negative effect of wild-type NS5A on NS5A-adapted subgenomic hepatitis C virus RNA replicon." Journal of General Virology 85, no. 7 (2004): 1867–75. http://dx.doi.org/10.1099/vir.0.80006-0.

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An efficient model is currently used to study hepatitis C virus (HCV) replication in cell culture. It involves transfection in Huh7, a hepatoma-derived cell line, of an antibiotic (neomycin) selectable HCV subgenomic replicon encoding the non-structural (NS) proteins from NS3 to NS5B. However, strong and sustained replication is achieved only on the appearance of adaptive mutations in viral proteins. The most effective of these adaptive mutations are concentrated mainly in NS5A, not only into the original Con1 but also in the recently established HCV-BK and HCV-H77 isolate-derived replicons. T

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8

Anraku, Itaru, Tracey J. Harvey, Richard Linedale, et al. "Kunjin Virus Replicon Vaccine Vectors Induce Protective CD8+ T-Cell Immunity." Journal of Virology 76, no. 8 (2002): 3791–99. http://dx.doi.org/10.1128/jvi.76.8.3791-3799.2002.

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ABSTRACT The ability of self-replicating RNA (replicon) vaccine vectors derived from the Australian flavivirus Kunjin (KUN) to induce protective αβ CD8+ T-cell responses was examined. KUN replicons encoding a model immunogen were delivered by three different vaccine modalities: (i) as naked RNA transcribed in vitro, (ii) as plasmid DNA constructed to allow in vivo transcription of replicon RNA by cellular RNA polymerase II (DNA based), and (iii) as replicon RNA encapsidated into virus-like particles. A single immunization with any of these KUN replicon vaccines induced CD8+ T-cell responses at

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9

Ali, Samir, Charles Pellerin, Daniel Lamarre, and George Kukolj. "Hepatitis C Virus Subgenomic Replicons in the Human Embryonic Kidney 293 Cell Line." Journal of Virology 78, no. 1 (2004): 491–501. http://dx.doi.org/10.1128/jvi.78.1.491-501.2004.

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ABSTRACT Hepatitis C virus (HCV) infects liver cells and its replication in other cells is incompletely defined. Human hepatoma Huh-7 cells harboring subgenomic HCV replicons were used in somatic cell fusion experiments with human embryonic kidney 293 cells as a means of examining the permissiveness of 293 cells for HCV subgenomic RNA replication. 293 cells were generally not permissive for replication of Huh-7 cell-adapted replicons. However, upon coculturing of the two cell lines, we selected rare replicon-containing cells, termed 293Rep cells, that resembled parental 293 cells. Direct metab

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10

Ammar, Ahmed M., Norhan K. Abd El-Aziz, Mohamed G. Aggour, et al. "A Newly Incompatibility F Replicon Allele (FIB81) in Extensively Drug-Resistant Escherichia coli Isolated from Diseased Broilers." International Journal of Molecular Sciences 25, no. 15 (2024): 8347. http://dx.doi.org/10.3390/ijms25158347.

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Multiple drug resistance (MDR) has gained pronounced attention among Enterobacterales. The transfer of multiple antimicrobial resistance genes, frequently carried on conjugative incompatibility F (IncF) plasmids and facilitating interspecies resistance transmission, has been linked to Salmonella spp. and E. coli in broilers. In Egypt, the growing resistance is exacerbated by the limited clinical efficacy of many antimicrobials. In this study, IncF groups were screened and characterized in drug-resistant Salmonella spp. and E. coli isolated from broilers. The antimicrobial resistance profile, P

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11

Erdmann, Maximilian, Peter A. C. Wing, Isobel Webb, et al. "A Novel Toolkit of SARS-CoV-2 Sub-Genomic Replicons for Efficient Antiviral Screening." Viruses 17, no. 5 (2025): 597. https://doi.org/10.3390/v17050597.

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SARS-CoV-2 is classified as a containment level 3 (CL3) pathogen, limiting research access and antiviral testing. To address this, we developed a non-infectious viral surrogate system using reverse genetics to generate sub-genomic replicons. These replicons contained the nsp1 mutations K164A and H165A and had the spike, membrane, ORF6, and ORF7a coding sequences replaced with various reporter and selectable marker genes. Replicons based on the ancestral Wuhan Hu-1 strain and the Delta variant of concern were replication-competent in multiple cell lines, as assessed by Renilla luciferase activi

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12

Gekeler, V., J. Epple, G. Kleymann, and H. Probst. "Selective and synchronous activation of early-S-phase replicons of Ehrlich ascites cells." Molecular and Cellular Biology 13, no. 8 (1993): 5020–33. http://dx.doi.org/10.1128/mcb.13.8.5020-5033.1993.

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Twelve-hour exposure of G1 Ehrlich ascites cells to controlled hypoxia (200 ppm of O2 at 1 bar) suppressed replicon initiation. Synchronous cycling, beginning with a normal S phase, was released by reoxygenation immediately. The addition of cycloheximide at reoxygenation largely resuppressed, after a short initial burst, succeeding replicon initiations. Alkaline sedimentation analysis of growing daughter strand DNA, DNA fiber autoradiography, and analysis of the newly formed DNA demonstrated that normal chain growth and DNA maturation (replicon termination) in the initially activated replicons

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13

Gekeler, V., J. Epple, G. Kleymann, and H. Probst. "Selective and synchronous activation of early-S-phase replicons of Ehrlich ascites cells." Molecular and Cellular Biology 13, no. 8 (1993): 5020–33. http://dx.doi.org/10.1128/mcb.13.8.5020.

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Twelve-hour exposure of G1 Ehrlich ascites cells to controlled hypoxia (200 ppm of O2 at 1 bar) suppressed replicon initiation. Synchronous cycling, beginning with a normal S phase, was released by reoxygenation immediately. The addition of cycloheximide at reoxygenation largely resuppressed, after a short initial burst, succeeding replicon initiations. Alkaline sedimentation analysis of growing daughter strand DNA, DNA fiber autoradiography, and analysis of the newly formed DNA demonstrated that normal chain growth and DNA maturation (replicon termination) in the initially activated replicons

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14

Targett-Adams, Paul, and John McLauchlan. "Development and characterization of a transient-replication assay for the genotype 2a hepatitis C virus subgenomic replicon." Journal of General Virology 86, no. 11 (2005): 3075–80. http://dx.doi.org/10.1099/vir.0.81334-0.

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Dicistronic, subgenomic hepatitis C virus (HCV) replicons were constructed containing sequences from JFH1, a genotype 2a strain, that also incorporated the firefly luciferase gene under the control of the HCV internal ribosome entry site element. Luciferase activity in Huh-7 cell extracts containing in vitro-transcribed subgenomic JFH1 RNA was monitored over a 72 h period to examine early stages of HCV replication in the absence of any selective pressure. Enzyme activities produced by the replicon were almost 200-fold greater than those generated from corresponding genotype 1b replicons and co

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15

Hu, Pingsheng, Xiaoming Chen, Lihong Huang, et al. "A highly pathogenic porcine reproductive and respiratory syndrome virus candidate vaccine based on Japanese encephalitis virus replicon system." PeerJ 5 (July 20, 2017): e3514. http://dx.doi.org/10.7717/peerj.3514.

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In the swine industry, porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease which causes heavy economic losses worldwide. Effective prevention and disease control is an important issue. In this study, we described the construction of a Japanese encephalitis virus (JEV) DNA-based replicon with a cytomegalovirus (CMV) promoter based on the genome of Japanese encephalitis live vaccine virus SA14-14-2, which is capable of offering a potentially novel way to develop and produce vaccines against a major pathogen of global health. This JEV DNA-based replicon contains a

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16

Friedhoff, Ronja, Ghada Elfayres, Natacha Mérindol, Isabel Desgagné-Penix, and Lionel Berthoux. "RNA replication-independent, DNA linearization-dependent expression of reporter genes from a SARS-CoV-2 replicon-encoding DNA in human cells." PLOS ONE 19, no. 8 (2024): e0300491. http://dx.doi.org/10.1371/journal.pone.0300491.

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Replicons, derived from RNA viruses, are genetic constructs retaining essential viral enzyme genes while lacking key structural protein genes. Upon introduction into cells, the genes carried by the replicon RNA are expressed, and the RNA self-replicates, yet viral particle production does not take place. Typically, RNA replicons are transcribed in vitro and are then electroporated in cells. However, it would be advantageous for the replicon to be generated in cells following DNA transfection instead of RNA. In this study, a bacterial artificial chromosome (BAC) DNA encoding a SARS-CoV-2 replic

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17

McCormick, Christopher J., Sophie Maucourant, Stephen Griffin, David J. Rowlands, and Mark Harris. "Tagging of NS5A expressed from a functional hepatitis C virus replicon." Journal of General Virology 87, no. 3 (2006): 635–40. http://dx.doi.org/10.1099/vir.0.81553-0.

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Knowledge of how hepatitis C virus (HCV) proteins associate with components of the host cell to form a functional replication complex is still limited. To address this issue, HCV replicon constructs were generated where either green fluorescent protein (GFP) or the Propionibacterium shermanii transcarboxylase domain (PSTCD) was introduced into the NS5A coding region. Insertion of both GFP and PSTCD was tolerated well, allowing formation of stable replicon-containing cell lines that contained viral protein and transcript levels that were comparable to those of an unmodified parental replicon. C

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18

Claus, Claudia, Wen-Pin Tzeng, Uwe G. Liebert, and Teryl K. Frey. "Rubella virus-induced superinfection exclusion studied in cells with persisting replicons." Journal of General Virology 88, no. 10 (2007): 2769–73. http://dx.doi.org/10.1099/vir.0.83092-0.

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For the first time, homologous superinfection exclusion was documented for rubella virus (RUB) by using Vero cells harbouring persisting RUB replicons. Infection with wild-type RUB was reduced by tenfold, whereas Sindbis virus infection was unaffected. Replication following infection with packaged replicons and transfection with replicon transcripts was also restricted in these cells, indicating that restriction occurred after penetration and entry. Translation of such ‘supertransfecting’ replicon transcripts was not impaired, but no accumulation of supertransfecting replicon RNA could be dete

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19

Ricardo-Lax, Inna, Joseph M. Luna, Tran Thi Nhu Thao, et al. "Replication and single-cycle delivery of SARS-CoV-2 replicons." Science 374, no. 6571 (2021): 1099–106. http://dx.doi.org/10.1126/science.abj8430.

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A tool to study SARS-CoV-2 Work with infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) requires high-level biocontainment facilities, making it important to develop safer molecular tools that can potentially be used under less stringent conditions. Self-replicating RNAs known as replicons have long been used to study pathogenic RNA viruses; however, developing replicons to study SARS-SoV-2 has been challenging because of its large genome. Ricardo-Lax et al . used a yeast-based system to construct SARS-CoV-2 replicons that cannot assemble infectious virus because they lack

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20

Hao, Weidong, Koleen J. Herlihy, Noelle Jie Zhang, et al. "Development of a Novel Dicistronic Reporter-Selectable Hepatitis C Virus Replicon Suitable for High-Throughput Inhibitor Screening." Antimicrobial Agents and Chemotherapy 51, no. 1 (2006): 95–102. http://dx.doi.org/10.1128/aac.01008-06.

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ABSTRACT Hepatitis C virus (HCV) research and drug discovery have been facilitated by the introduction of cell lines with self-replicating subgenomic HCV replicons. Early attempts to carry out robust, high-throughput screens (HTS) using HCV replicons have met with limited success. Specifically, selectable replicons have required laborious reverse transcription-PCR quantitation, and reporter replicons have generated low signal-to-noise ratios. In this study, we constructed a dicistronic single reporter (DSR)-selectable HCV replicon that contained a humanized Renilla luciferase (hRLuc) gene sepa

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Lam, Angela M. I., and David N. Frick. "Hepatitis C Virus Subgenomic Replicon Requires an Active NS3 RNA Helicase." Journal of Virology 80, no. 1 (2006): 404–11. http://dx.doi.org/10.1128/jvi.80.1.404-411.2006.

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ABSTRACT Mutations were introduced into the NS3 helicase region of a hepatitis C virus (HCV) Con1 subgenomic replicon to ascertain the role of the helicase in viral replication. One new replicon lacked two-thirds of the NS3 helicase (Δhel), and six others contained one of the following six amino acid substitutions in NS3: R393A, F438A, T450I, E493K, W501A, and W501F. It has been previously reported that purified R393A, F438A, and W501A HCV helicase proteins do not unwind RNA but unwind DNA, bind RNA, and hydrolyze ATP. On the other hand, previous data suggest that a W501F protein retains most

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Shih, I.-hung, Inge Vliegen, Betty Peng, et al. "Mechanistic Characterization of GS-9190 (Tegobuvir), a Novel Nonnucleoside Inhibitor of Hepatitis C Virus NS5B Polymerase." Antimicrobial Agents and Chemotherapy 55, no. 9 (2011): 4196–203. http://dx.doi.org/10.1128/aac.00307-11.

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ABSTRACTGS-9190 (Tegobuvir) is a novel imidazopyridine inhibitor of hepatitis C virus (HCV) RNA replicationin vitroand has demonstrated potent antiviral activity in patients chronically infected with genotype 1 (GT1) HCV. GS-9190 exhibits reduced activity against GT2a (JFH1) subgenomic replicons and GT2a (J6/JFH1) infectious virus, suggesting that the compound's mechanism of action involves a genotype-specific viral component. To further investigate the GS-9190 mechanism of action, we utilized the susceptibility differences between GT1b and GT2a by constructing a series of replicon chimeras wh

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23

Robinson, Margaret, Huiling Yang, Siu-Chi Sun, et al. "Novel Hepatitis C Virus Reporter Replicon Cell Lines Enable Efficient Antiviral Screening against Genotype 1a." Antimicrobial Agents and Chemotherapy 54, no. 8 (2010): 3099–106. http://dx.doi.org/10.1128/aac.00289-10.

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ABSTRACT The hepatitis C virus (HCV) subgenomic replicon is the primary tool for evaluating the activity of anti-HCV compounds in drug discovery research. Despite the prevalence of HCV genotype 1a (∼70% of U.S. HCV patients), few genotype 1a reporter replicon cell lines have been described; this is presumably due to the low replication capacity of such constructs in available Huh-7 cells. In this report, we describe the selection of highly permissive Huh-7 cell lines that support robust replication of genotype 1a subgenomic replicons harboring luciferase reporter genes. These novel cell lines

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Gu, Baohua, Adam T. Gates, Olaf Isken, Sven-Erik Behrens, and Robert T. Sarisky. "Replication Studies Using Genotype 1a Subgenomic Hepatitis C Virus Replicons." Journal of Virology 77, no. 9 (2003): 5352–59. http://dx.doi.org/10.1128/jvi.77.9.5352-5359.2003.

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ABSTRACT Recently, cell-based replicon systems for hepatitis C virus (HCV), in which the nonstructural proteins stably replicate subgenomic viral RNA in Huh7 cells, were developed. To date, one limitation of using these replicon systems to advance drug discovery is the inability of other genotypic derivatives, beyond those of two distinct strains of genotype 1b (HCV-N and Con1), to stably replicate in Huh7 cells. In this report, we evaluated a series of replicon genotype 1a-1b chimeras, as well as a complete genotype 1a replicon clone. A subgenomic replicon construct containing only type 1a se

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Perri, Silvia, David A. Driver, Jason P. Gardner, et al. "Replicon Vectors Derived from Sindbis Virus and Semliki Forest Virus That Establish Persistent Replication in Host Cells." Journal of Virology 74, no. 20 (2000): 9802–7. http://dx.doi.org/10.1128/jvi.74.20.9802-9807.2000.

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ABSTRACT Alphavirus replicon vectors are well suited for applications where transient, high-level expression of a heterologous gene is required. Replicon vector expression in cells leads to inhibition of host macromolecular synthesis, culminating in eventual cell death by an apoptotic mechanism. For many applications, including gene expression studies in cultured cells, a longer duration of transgene expression without resulting cytopathic effects is useful. Recently, noncytopathic Sindbis virus (SIN) variants were isolated in BHK cells, and the mutations responsible were mapped to the proteas

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Korba, Brent E., Menashe Elazar, Ping Lui, Jean-François Rossignol, and Jeffrey S. Glenn. "Potential for Hepatitis C Virus Resistance to Nitazoxanide or Tizoxanide." Antimicrobial Agents and Chemotherapy 52, no. 11 (2008): 4069–71. http://dx.doi.org/10.1128/aac.00078-08.

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ABSTRACT Nitazoxanide and its primary metabolite, tizoxanide, inhibit hepatitis C virus (HCV) replication in HCV replicon systems. To study the potential for resistance, we subjected Huh7 cells harboring HCV replicons to serial passage in 250 μM G418 and increasing concentrations of nitazoxanide or tizoxanide. Passage of the replicon-containing cell lines in either compound resulted in increases in the 50% effective concentrations (EC50s) (7- to 13-fold), EC90s (14- to 36-fold), and 50% cytotoxic concentrations (2- to 4-fold) of both compounds. Serial passage in either compound did not alter t

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Flint, Mike, Stanley Mullen, Anne M. Deatly, et al. "Selection and Characterization of Hepatitis C Virus Replicons Dually Resistant to the Polymerase and Protease Inhibitors HCV-796 and Boceprevir (SCH 503034)." Antimicrobial Agents and Chemotherapy 53, no. 2 (2008): 401–11. http://dx.doi.org/10.1128/aac.01081-08.

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ABSTRACT HCV-796 is a nonnucleoside inhibitor of the hepatitis C virus (HCV) nonstructural protein 5B (NS5B) polymerase, and boceprevir is an inhibitor of the NS3 serine protease. The emergence of replicon variants resistant to the combination of HCV-796 and boceprevir was evaluated. Combining the inhibitors greatly reduced the frequency with which resistant colonies arose; however, some resistant replicon cells could be isolated by the use of low inhibitor concentrations. These replicons were approximately 1,000-fold less susceptible to HCV-796 and 9-fold less susceptible to boceprevir. They

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Cheng, Wen-Fang, Chien-Fu Hung, Chee-Yin Chai, et al. "Enhancement of Sindbis Virus Self-Replicating RNA Vaccine Potency by Linkage of Herpes Simplex Virus Type 1 VP22 Protein to Antigen." Journal of Virology 75, no. 5 (2001): 2368–76. http://dx.doi.org/10.1128/jvi.75.5.2368-2376.2001.

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ABSTRACT Recently, self-replicating and self-limiting RNA vaccines (RNA replicons) have emerged as an important form of nucleic acid vaccines. Self-replicating RNA eventually causes lysis of transfected cells and does not raise the concern associated with naked DNA vaccines of integration into the host genome. This is particularly important for development of vaccines targeting proteins that are potentially oncogenic. However, the potency of RNA replicons is significantly limited by their lack of intrinsic ability to spread in vivo. The herpes simplex virus type 1 protein VP22 has demonstrated

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Reap, Elizabeth A., Sergey A. Dryga, John Morris, et al. "Cellular and Humoral Immune Responses to Alphavirus Replicon Vaccines Expressing Cytomegalovirus pp65, IE1, and gB Proteins." Clinical and Vaccine Immunology 14, no. 6 (2007): 748–55. http://dx.doi.org/10.1128/cvi.00037-07.

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ABSTRACT Development of vaccines against cytomegalovirus (CMV) is an important public health priority. We used a propagation-defective, single-cycle RNA replicon vector system derived from an attenuated strain of an alphavirus, Venezuelan equine encephalitis virus, to produce virus-like replicon particles (VRP) expressing various combinations of pp65, IE1, or gB proteins of human CMV. Protein expression in VRP-infected cells was highest with single-promoter replicons expressing pp65, IE1, a pp65/IE1 fusion protein, or the extracellular domain of gB and with double-promoter replicons expressing

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Gehrke, Rainer, Michael Ecker, Stephan W. Aberle, Steven L. Allison, Franz X. Heinz, and Christian W. Mandl. "Incorporation of Tick-Borne Encephalitis Virus Replicons into Virus-Like Particles by a Packaging Cell Line." Journal of Virology 77, no. 16 (2003): 8924–33. http://dx.doi.org/10.1128/jvi.77.16.8924-8933.2003.

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ABSTRACT RNA replicons derived from flavivirus genomes show considerable potential as gene transfer and immunization vectors. A convenient and efficient encapsidation system is an important prerequisite for the practical application of such vectors. In this work, tick-borne encephalitis (TBE) virus replicons and an appropriate packaging cell line were constructed and characterized. A stable CHO cell line constitutively expressing the two surface proteins prM/M and E (named CHO-ME cells) was generated and shown to efficiently export mature recombinant subviral particles (RSPs). When replicon Nd

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Tzeng, Wen-Pin, and Teryl K. Frey. "Complementation of a Deletion in the Rubella Virus P150 Nonstructural Protein by the Viral Capsid Protein." Journal of Virology 77, no. 17 (2003): 9502–10. http://dx.doi.org/10.1128/jvi.77.17.9502-9510.2003.

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ABSTRACT Rubella virus (RUB) replicons with an in-frame deletion of 507 nucleotides between two NotI sites in the P150 nonstructural protein (ΔNotI) do not replicate (as detected by expression of a reporter gene encoded by the replicon) but can be amplified by wild-type helper virus (Tzeng et al., Virology 289:63-73, 2001). Surprisingly, virus with ΔNotI was viable, and it was hypothesized that this was due to complementation of the NotI deletion by one of the virion structural protein genes. Introduction of the capsid (C) protein gene into ΔNotI-containing replicons as an in-frame fusion with

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Queiroz, Sabrina R. A., Andréa N. M. R. Silva, Jefferson J. S. Santos, Ernesto T. A. Marques Jr, Giovani R. Bertani, and Laura H. V. G. Gil. "Construction of yellow fever virus subgenomic replicons by yeast-based homologous recombination cloning technique." Anais da Academia Brasileira de Ciências 85, no. 1 (2013): 159–68. http://dx.doi.org/10.1590/s0001-37652013005000008.

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RNA replicon derived from Flavivirus genome is a valuable tool for studying viral replication independent of virion assembly and maturation, besides being a great potencial for heterologous gene expression. In this study we described the construction of subgenomic replicons of yellow fever virus by yeast-based homologous recombination technique. The plasmid containing the yellow fever 17D strain replicon (pBSC-repYFV-17D), previously characterized, was handled to heterologous expression of the green fluorescent protein (repYFV-17D-GFP) and firefly luciferase (repYFV-17D-Luc) reporter genes. Bo

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Brown, Ashley N., James J. McSharry, Jonathan R. Adams, et al. "Pharmacodynamic Analysis of a Serine Protease Inhibitor, MK-4519, against Hepatitis C Virus Using a NovelIn VitroPharmacodynamic System." Antimicrobial Agents and Chemotherapy 56, no. 3 (2011): 1170–81. http://dx.doi.org/10.1128/aac.05383-11.

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ABSTRACTThe development of new antiviral compounds active against hepatitis C virus (HCV) has surged in recent years. In order for these new compounds to be efficacious in humans, optimal dosage regimens for each compound must be elucidated. We have developed a novelin vitropharmacokinetic/pharmacodynamic system, the BelloCell system, to identify optimal dosage regimens for anti-HCV compounds. In these experiments, genotype 1b HCV replicon-bearing cells (2209-23 cells) were inoculated onto carrier flakes in BelloCell bottles and treated with MK-4519, a serine protease inhibitor. Our dose-rangi

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Harvey, Tracey J., Itaru Anraku, Richard Linedale, et al. "Kunjin Virus Replicon Vectors for Human Immunodeficiency Virus Vaccine Development." Journal of Virology 77, no. 14 (2003): 7796–803. http://dx.doi.org/10.1128/jvi.77.14.7796-7803.2003.

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ABSTRACT We have previously demonstrated the ability of the vaccine vectors based on replicon RNA of the Australian flavivirus Kunjin (KUN) to induce protective antiviral and anticancer CD8+ T-cell responses using murine polyepitope as a model immunogen (I. Anraku, T. J. Harvey, R. Linedale, J. Gardner, D. Harrich, A. Suhrbier, and A. A. Khromykh, J. Virol. 76:3791-3799, 2002). Here we showed that immunization of BALB/c mice with KUN replicons encoding HIV-1 Gag antigen resulted in induction of both Gag-specific antibody and protective Gag-specific CD8+ T-cell responses. Two immunizations with

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Johnson, Timothy J., Yvonne M. Wannemuehler, Sara J. Johnson, et al. "Plasmid Replicon Typing of Commensal and Pathogenic Escherichia coli Isolates." Applied and Environmental Microbiology 73, no. 6 (2007): 1976–83. http://dx.doi.org/10.1128/aem.02171-06.

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ABSTRACT Despite the critical role of plasmids in horizontal gene transfer, few studies have characterized plasmid relatedness among different bacterial populations. Recently, a multiplex PCR replicon typing protocol was developed for classification of plasmids occurring in members of the Enterobacteriaceae. Here, a simplified version of this replicon typing procedure which requires only three multiplex panels to identify 18 plasmid replicons is described. This method was used to screen 1,015 Escherichia coli isolates of avian, human, and poultry meat origin for plasmid replicon types. Additio

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Lam, Angela M., Christine Espiritu, Shalini Bansal, et al. "Genotype and Subtype Profiling of PSI-7977 as a Nucleotide Inhibitor of Hepatitis C Virus." Antimicrobial Agents and Chemotherapy 56, no. 6 (2012): 3359–68. http://dx.doi.org/10.1128/aac.00054-12.

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ABSTRACTPSI-7977, a prodrug of 2′-F-2′-C-methyluridine monophosphate, is the purified diastereoisomer of PSI-7851 and is currently being investigated in phase 3 clinical trials for the treatment of hepatitis C. In this study, we profiled the activity of PSI-7977 and its ability to select for resistance using a number of different replicon cells. Results showed that PSI-7977 was active against genotype (GT) 1a, 1b, and 2a (strain JFH-1) replicons and chimeric replicons containing GT 2a (strain J6), 2b, and 3a NS5B polymerase. Cross-resistance studies using GT 1b replicons confirmed that the S28

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Le Pogam, Sophie, Hyunsoon Kang, Seth F. Harris, et al. "Selection and Characterization of Replicon Variants Dually Resistant to Thumb- and Palm-Binding Nonnucleoside Polymerase Inhibitors of the Hepatitis C Virus." Journal of Virology 80, no. 12 (2006): 6146–54. http://dx.doi.org/10.1128/jvi.02628-05.

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ABSTRACT Multiple nonnucleoside inhibitor binding sites have been identified within the hepatitis C virus (HCV) polymerase, including in the palm and thumb domains. After a single treatment with a thumb site inhibitor (thiophene-2-carboxylic acid NNI-1), resistant HCV replicon variants emerged that contained mutations at residues Leu419, Met423, and Ile482 in the polymerase thumb domain. Binding studies using wild-type (WT) and mutant enzymes and structure-based modeling showed that the mechanism of resistance is through the reduced binding of the inhibitor to the mutant enzymes. Combined trea

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Tocchetti, Arianna, Gloria Galimberti, Gianni Dehò, and Daniela Ghisotti. "Characterization of the oriI andoriII Origins of Replication in Phage-Plasmid P4." Journal of Virology 73, no. 9 (1999): 7308–16. http://dx.doi.org/10.1128/jvi.73.9.7308-7316.1999.

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ABSTRACT In the Escherichia coli phage-plasmid P4, two partially overlapping replicons with bipartite ori sites coexist. The essential components of the oriI replicon are the α andcnr genes and the ori1 and crrsites; the oriII replicon is composed of the α gene, with the internal ori2 site, and the crr region. The P4 α protein has primase and helicase activities and specifically binds type I iterons, present in ori1 and crr. Using a complementation test for plasmid replication, we demonstrated that the two replicons depend on both the primase and helicase activities of the α protein. Moreover,

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Ivanova, Lidia, Sondra Schlesinger, and Paul D. Olivo. "Regulated Expression of a Sindbis Virus Replicon by Herpesvirus Promoters." Journal of Virology 73, no. 3 (1999): 1998–2005. http://dx.doi.org/10.1128/jvi.73.3.1998-2005.1999.

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ABSTRACT We describe the use of herpesvirus promoters to regulate the expression of a Sindbis virus replicon (SINrep/LacZ). We isolated cell lines that contain the cDNA of SINrep/LacZ under the control of a promoter from a herpesvirus early gene which requires regulatory proteins encoded by immediate-early genes for expression. Wild-type Sindbis virus and replicons derived from this virus cause death of most vertebrate cells, but the cells discussed here grew normally and expressed the replicon and β-galactosidase only after infection with a herpesvirus. Vero cell lines in which the expression

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Tzeng, Wen-Pin, Jason D. Matthews, and Teryl K. Frey. "Analysis of Rubella Virus Capsid Protein-Mediated Enhancement of Replicon Replication and Mutant Rescue." Journal of Virology 80, no. 8 (2006): 3966–74. http://dx.doi.org/10.1128/jvi.80.8.3966-3974.2006.

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ABSTRACT The rubella virus capsid protein (C) has been shown to complement a lethal deletion (termed ΔNotI) in P150 replicase protein. To investigate this phenomenon, we generated two lines of Vero cells that stably expressed either C (C-Vero cells) or C lacking the eight N-terminal residues (CΔ8-Vero cells), a construct previously shown to be unable to complement ΔNotI. In C-Vero cells but not Vero or CΔ8-Vero cells, replication of a wild-type (wt) replicon expressing the green fluorescent protein (GFP) reporter gene (RUBrep/GFP) was enhanced, and replication of a replicon with ΔNotI (RUBrep/

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Yang, Feng, Jason M. Robotham, Heather B. Nelson, Andre Irsigler, Rachael Kenworthy, and Hengli Tang. "Cyclophilin A Is an Essential Cofactor for Hepatitis C Virus Infection and the Principal Mediator of Cyclosporine Resistance In Vitro." Journal of Virology 82, no. 11 (2008): 5269–78. http://dx.doi.org/10.1128/jvi.02614-07.

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ABSTRACT Cyclosporine (CsA) and its derivatives potently suppress hepatitis C virus (HCV) replication. Recently, CsA-resistant HCV replicons have been identified in vitro. We examined the dependence of the wild-type and CsA-resistant replicons on various cyclophilins for replication. A strong correlation between CsA resistance and reduced dependency on cyclophilin A (CyPA) for replication was identified. Silencing of CyPB or CyPC expression had no significant effect on replication, whereas various forms of small interfering RNA (siRNA) directed at CyPA inhibited HCV replication of wild-type bu

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Evans, Matthew J., Charles M. Rice, and Stephen P. Goff. "Genetic Interactions between Hepatitis C Virus Replicons." Journal of Virology 78, no. 21 (2004): 12085–89. http://dx.doi.org/10.1128/jvi.78.21.12085-12089.2004.

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ABSTRACT To investigate interactions between hepatitis C virus (HCV) RNA replication complexes, a system was developed to simultaneously select different HCV subgenomic replicons within the same cell. Transcomplementation of defective replicons was not observed, suggesting an isolated and independent nature of the HCV RNA replication complex. In contrast, a high level of competition between replicons was observed, such that the presence and increased fitness of one replicon reduced the capacity of a second one to stably replicate. These results suggest that at least one factor in Huh7 cells re

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Bartosik, Dariusz, Michal Szymanik, and Edyta Wysocka. "Identification of the Partitioning Site within therepABC-Type Replicon of the Composite Paracoccus versutus Plasmid pTAV1." Journal of Bacteriology 183, no. 21 (2001): 6234–43. http://dx.doi.org/10.1128/jb.183.21.6234-6243.2001.

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ABSTRACT The replicator region of composite plasmid pTAV1 ofParacoccus versutus (included in mini-replicon pTAV320) belongs to the family of repABC replicons commonly found in plasmids harbored by Agrobacterium andRhizobium spp. The repABC replicons encode three genes clustered in an operon, which are involved in partitioning (repA and repB) and replication (repC). In order to localize the partitioning site of pTAV320, the two identified incompatibility determinants of this mini-replicon (inc1, located in the intergenic sequence between repB andrepC; and inc2, situated downstream of the repC g

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Hughes, Mair, Sarah Gretton, Holly Shelton, et al. "A Conserved Proline between Domains II and III of Hepatitis C Virus NS5A Influences both RNA Replication and Virus Assembly." Journal of Virology 83, no. 20 (2009): 10788–96. http://dx.doi.org/10.1128/jvi.02406-08.

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ABSTRACT We previously demonstrated that two closely spaced polyproline motifs, with the consensus sequence Pro-X-X-Pro-X-Lys/Arg, located between residues 343 to 356 of NS5A, mediated interactions with cellular SH3 domains. The N-terminal motif (termed PP2.1) is only conserved in genotype 1 isolates, whereas the C-terminal motif (PP2.2) is conserved throughout all hepatitis C virus (HCV) isolates, although this motif was shown to be dispensable for replication of the genotype 1b subgenomic replicon. In order to investigate the potential role of these motifs in the viral life cycle, we have un

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Sawicki, Dorothea L., Silvia Perri, John M. Polo, and Stanley G. Sawicki. "Role for nsP2 Proteins in the Cessation of Alphavirus Minus-Strand Synthesis by Host Cells." Journal of Virology 80, no. 1 (2006): 360–71. http://dx.doi.org/10.1128/jvi.80.1.360-371.2006.

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ABSTRACT In order to establish nonlytic persistent infections (PI) of BHK cells, replicons derived from Sindbis (SIN) and Semliki Forest (SFV) viruses have mutations in nsP2. Five different nsP2 PI replicons were compared to wild-type (wt) SIN, SFV, and wt nsPs SIN replicons. Replicon PI BHK21 cells had viral RNA synthesis rates that were less than 5% of those of the wt virus and ∼10% or less of those of SIN wt replicon-infected cells, and, in contrast to wt virus and replicons containing wt nsP2, all showed a phenotype of continuous minus-strand synthesis and of unstable, mature replication/t

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Johansen, Lisa K., and Casey D. Morrow. "Inherent Instability of Poliovirus Genomes Containing Two Internal Ribosome Entry Site (IRES) Elements Supports a Role for the IRES in Encapsidation." Journal of Virology 74, no. 18 (2000): 8335–42. http://dx.doi.org/10.1128/jvi.74.18.8335-8342.2000.

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ABSTRACT Previous studies have described poliovirus genomes in which the internal ribosome entry (IRES) for encephalomyocarditis virus (EMCV) is positioned between the P1 and P2-P3 open reading frames of the poliovirus genome. Although these dicistronic poliovirus genomes were replication competent, most exhibited evidence of genetic instability, and the EMCV IRES was deleted upon serial passage. One possible reason for instability of the genome is that the dicistronic genome was at least 108% larger than the wild-type poliovirus genome, which could reduce the efficiency of encapsidation. To a

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Berezney, Ronald, Dharani D. Dubey, and Joel A. Huberman. "Heterogeneity of eukaryotic replicons, replicon clusters, and replication foci." Chromosoma 108, no. 8 (2000): 471–84. http://dx.doi.org/10.1007/s004120050399.

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Herlihy, Koleen J., Joanne P. Graham, Robert Kumpf, Amy K. Patick, Rohit Duggal, and Stephanie T. Shi. "Development of Intergenotypic Chimeric Replicons To Determine the Broad-Spectrum Antiviral Activities of Hepatitis C Virus Polymerase Inhibitors." Antimicrobial Agents and Chemotherapy 52, no. 10 (2008): 3523–31. http://dx.doi.org/10.1128/aac.00533-08.

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ABSTRACT To address the need for broad-spectrum antiviral activity characterization of hepatitis C virus (HCV) polymerase inhibitors, we created a panel of intergenotypic chimeric replicons containing nonstructural (NS) protein NS5B sequences from genotype 2b (GT2b), GT3a, GT4a, GT5a, and GT6a HCV isolates. Viral RNA extracted from non-GT1 HCV patient plasma was subjected to reverse transcription. The NS5B region was amplified by nested PCR and introduced into the corresponding region of the GT1b (Con-1) subgenomic reporter replicon by Splicing by Overlap Extension (SOEing) PCR. Stable cell li

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Risager, Peter Christian, Ulrik Fahnøe, Maria Gullberg, Thomas Bruun Rasmussen, and Graham J. Belsham. "Analysis of classical swine fever virus RNA replication determinants using replicons." Journal of General Virology 94, no. 8 (2013): 1739–48. http://dx.doi.org/10.1099/vir.0.052688-0.

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Self-replicating RNAs (replicons), with or without reporter gene sequences, derived from the genome of the Paderborn strain of classical swine fever virus (CSFV) have been produced. The full-length viral cDNA, propagated within a bacterial artificial chromosome, was modified by targeted recombination within Escherichia coli. RNA transcripts were produced in vitro and introduced into cells by electroporation. The translation and replication of the replicon RNAs could be followed by the accumulation of luciferase (from Renilla reniformis or Gaussia princeps) protein expression (where appropriate

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Yang, Yang, Jiayang Zheng, Yafang Lin, et al. "Development of a Noninfectious Japanese Encephalitis Virus Replicon for Antiviral Drug Screening and Gene Function Studies." Viruses 17, no. 6 (2025): 759. https://doi.org/10.3390/v17060759.

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Viral replicons are efficient tools to understand the mechanisms of viral replication and screen antiviral drugs. In this study, a viral-cDNA-based replicon of Japanese encephalitis virus (JEV), which is the causative agent of Japanese encephalitis, was constructed by replacing the viral structural proteins with a green fluorescent protein (JEV-GFP replicon). The resulting JEV-GFP replicon was used as a tool to screen antiviral drugs targeting JEV nonstructural proteins, and the five compounds JNJ-A07, HZ-1157, NITD-2, quinine, and NITD008 were obtained, which significantly inhibited the repli

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