A subclone of HuH-7 with enhanced intracellular hepatitis C virus production and evasion of virus related-cell cycle arrest
Hepatitis C virus (HCV) cell culture system with JFH-1 strain and HuH-7 cells enabled us to produce infectious HCV particles in vitro, and such system is useful to explore the anti-HCV compounds and to develop the vaccine against HCV. In the present study, we describe the derivation of a cell line t...
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| Veröffentlicht in: | PloS one 2012-12, Vol.7 (12), p.e52697-e52697 |
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| creator | Murayama, Asako Sugiyama, Nao Yoshimura, Seiko Ishihara-Sugano, Mitsuko Masaki, Takahiro Kim, Sulyi Wakita, Takaji Mishiro, Shunji Kato, Takanobu |
| description | Hepatitis C virus (HCV) cell culture system with JFH-1 strain and HuH-7 cells enabled us to produce infectious HCV particles in vitro, and such system is useful to explore the anti-HCV compounds and to develop the vaccine against HCV. In the present study, we describe the derivation of a cell line that permits improved production of HCV particles. Specifically, we characterized several subclones that were isolated from the original HuH-7 cell line by limiting dilution. These HuH-7 subclones displayed a notable range of HCV production levels following transfection by full-genome JFH-1 RNA. Among these subclones, HuH-7T1 produced HCV more efficiently than other subclones and Huh-7.5.1 that is known to be highly permissive for HCV replication. Upon transfection with full-genome RNA, HCV production was increased ten-fold in HuH-7T1 compared to Huh-7.5.1. This increase in viral production correlated with increased efficiency of intracellular infectious virus production. Furthermore, HCV replication did not induce cell cycle arrest in HuH-7T1, whereas it did in Huh-7.5.1. Consequently, the use of HuH-7T1 as host cells could provide increased population of HCV-positive cells and elevated viral titer. In conclusion, we isolated a HuH-7 subclone, HuH-7T1, that supports efficient HCV production. High efficiency of intracellular infectious virus production and evasion of cell cycle arrest were important for this phenotype. We expect that the use of this cell line will facilitate analysis of the underlying mechanisms for HCV particle assembly and the cell cycle arrest caused by HCV. |
| doi_str_mv | 10.1371/journal.pone.0052697 |
| format | Article |
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In the present study, we describe the derivation of a cell line that permits improved production of HCV particles. Specifically, we characterized several subclones that were isolated from the original HuH-7 cell line by limiting dilution. These HuH-7 subclones displayed a notable range of HCV production levels following transfection by full-genome JFH-1 RNA. Among these subclones, HuH-7T1 produced HCV more efficiently than other subclones and Huh-7.5.1 that is known to be highly permissive for HCV replication. Upon transfection with full-genome RNA, HCV production was increased ten-fold in HuH-7T1 compared to Huh-7.5.1. This increase in viral production correlated with increased efficiency of intracellular infectious virus production. Furthermore, HCV replication did not induce cell cycle arrest in HuH-7T1, whereas it did in Huh-7.5.1. Consequently, the use of HuH-7T1 as host cells could provide increased population of HCV-positive cells and elevated viral titer. In conclusion, we isolated a HuH-7 subclone, HuH-7T1, that supports efficient HCV production. High efficiency of intracellular infectious virus production and evasion of cell cycle arrest were important for this phenotype. We expect that the use of this cell line will facilitate analysis of the underlying mechanisms for HCV particle assembly and the cell cycle arrest caused by HCV.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0052697</identifier><identifier>PMID: 23285155</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Biology ; Cell culture ; Cell cycle ; Cell Cycle Checkpoints ; Cell growth ; Cell Line ; Cloning ; Dilution ; Gene Expression Regulation ; Genomes ; Genotype ; Genotype & phenotype ; Hepacivirus - physiology ; Hepatitis ; Hepatitis C ; Hepatitis C virus ; Humans ; Infection ; Infectious diseases ; Intracellular ; Kinases ; Laboratories ; Liver cancer ; Medicine ; Proteins ; R&D ; Replication ; Research & development ; Ribonucleic acid ; RNA ; RNA polymerase ; Transfection ; Vaccines ; Viral Load ; Virology ; Virus Replication ; Viruses</subject><ispartof>PloS one, 2012-12, Vol.7 (12), p.e52697-e52697</ispartof><rights>COPYRIGHT 2012 Public Library of Science</rights><rights>2012 Murayama et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2012 Murayama et al 2012 Murayama et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-c7058329d2cf2b43f75bf86fe95d801c8e8f262167986d14cbfe33cbc83ddf503</citedby><cites>FETCH-LOGICAL-c692t-c7058329d2cf2b43f75bf86fe95d801c8e8f262167986d14cbfe33cbc83ddf503</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3527576/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3527576/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79342,79343</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23285155$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Murayama, Asako</creatorcontrib><creatorcontrib>Sugiyama, Nao</creatorcontrib><creatorcontrib>Yoshimura, Seiko</creatorcontrib><creatorcontrib>Ishihara-Sugano, Mitsuko</creatorcontrib><creatorcontrib>Masaki, Takahiro</creatorcontrib><creatorcontrib>Kim, Sulyi</creatorcontrib><creatorcontrib>Wakita, Takaji</creatorcontrib><creatorcontrib>Mishiro, Shunji</creatorcontrib><creatorcontrib>Kato, Takanobu</creatorcontrib><title>A subclone of HuH-7 with enhanced intracellular hepatitis C virus production and evasion of virus related-cell cycle arrest</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Hepatitis C virus (HCV) cell culture system with JFH-1 strain and HuH-7 cells enabled us to produce infectious HCV particles in vitro, and such system is useful to explore the anti-HCV compounds and to develop the vaccine against HCV. 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In conclusion, we isolated a HuH-7 subclone, HuH-7T1, that supports efficient HCV production. High efficiency of intracellular infectious virus production and evasion of cell cycle arrest were important for this phenotype. We expect that the use of this cell line will facilitate analysis of the underlying mechanisms for HCV particle assembly and the cell cycle arrest caused by HCV.</description><subject>Analysis</subject><subject>Biology</subject><subject>Cell culture</subject><subject>Cell cycle</subject><subject>Cell Cycle Checkpoints</subject><subject>Cell growth</subject><subject>Cell Line</subject><subject>Cloning</subject><subject>Dilution</subject><subject>Gene Expression Regulation</subject><subject>Genomes</subject><subject>Genotype</subject><subject>Genotype & phenotype</subject><subject>Hepacivirus - physiology</subject><subject>Hepatitis</subject><subject>Hepatitis C</subject><subject>Hepatitis C virus</subject><subject>Humans</subject><subject>Infection</subject><subject>Infectious diseases</subject><subject>Intracellular</subject><subject>Kinases</subject><subject>Laboratories</subject><subject>Liver 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subclone of HuH-7 with enhanced intracellular hepatitis C virus production and evasion of virus related-cell cycle arrest</title><author>Murayama, Asako ; Sugiyama, Nao ; Yoshimura, Seiko ; Ishihara-Sugano, Mitsuko ; Masaki, Takahiro ; Kim, Sulyi ; Wakita, Takaji ; Mishiro, Shunji ; Kato, Takanobu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-c7058329d2cf2b43f75bf86fe95d801c8e8f262167986d14cbfe33cbc83ddf503</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Analysis</topic><topic>Biology</topic><topic>Cell culture</topic><topic>Cell cycle</topic><topic>Cell Cycle Checkpoints</topic><topic>Cell growth</topic><topic>Cell Line</topic><topic>Cloning</topic><topic>Dilution</topic><topic>Gene Expression Regulation</topic><topic>Genomes</topic><topic>Genotype</topic><topic>Genotype & phenotype</topic><topic>Hepacivirus - 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Nao</au><au>Yoshimura, Seiko</au><au>Ishihara-Sugano, Mitsuko</au><au>Masaki, Takahiro</au><au>Kim, Sulyi</au><au>Wakita, Takaji</au><au>Mishiro, Shunji</au><au>Kato, Takanobu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A subclone of HuH-7 with enhanced intracellular hepatitis C virus production and evasion of virus related-cell cycle arrest</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2012-12-20</date><risdate>2012</risdate><volume>7</volume><issue>12</issue><spage>e52697</spage><epage>e52697</epage><pages>e52697-e52697</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Hepatitis C virus (HCV) cell culture system with JFH-1 strain and HuH-7 cells enabled us to produce infectious HCV particles in vitro, and such system is useful to explore the anti-HCV compounds and to develop the vaccine against HCV. In the present study, we describe the derivation of a cell line that permits improved production of HCV particles. Specifically, we characterized several subclones that were isolated from the original HuH-7 cell line by limiting dilution. These HuH-7 subclones displayed a notable range of HCV production levels following transfection by full-genome JFH-1 RNA. Among these subclones, HuH-7T1 produced HCV more efficiently than other subclones and Huh-7.5.1 that is known to be highly permissive for HCV replication. Upon transfection with full-genome RNA, HCV production was increased ten-fold in HuH-7T1 compared to Huh-7.5.1. This increase in viral production correlated with increased efficiency of intracellular infectious virus production. Furthermore, HCV replication did not induce cell cycle arrest in HuH-7T1, whereas it did in Huh-7.5.1. Consequently, the use of HuH-7T1 as host cells could provide increased population of HCV-positive cells and elevated viral titer. In conclusion, we isolated a HuH-7 subclone, HuH-7T1, that supports efficient HCV production. High efficiency of intracellular infectious virus production and evasion of cell cycle arrest were important for this phenotype. We expect that the use of this cell line will facilitate analysis of the underlying mechanisms for HCV particle assembly and the cell cycle arrest caused by HCV.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23285155</pmid><doi>10.1371/journal.pone.0052697</doi><tpages>e52697</tpages><oa>free_for_read</oa></addata></record> |
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| source | Public Library of Science (PLoS) Journals Open Access; MEDLINE; DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Analysis Biology Cell culture Cell cycle Cell Cycle Checkpoints Cell growth Cell Line Cloning Dilution Gene Expression Regulation Genomes Genotype Genotype & phenotype Hepacivirus - physiology Hepatitis Hepatitis C Hepatitis C virus Humans Infection Infectious diseases Intracellular Kinases Laboratories Liver cancer Medicine Proteins R&D Replication Research & development Ribonucleic acid RNA RNA polymerase Transfection Vaccines Viral Load Virology Virus Replication Viruses |
| title | A subclone of HuH-7 with enhanced intracellular hepatitis C virus production and evasion of virus related-cell cycle arrest |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-04T21%3A26%3A28IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=A%20subclone%20of%20HuH-7%20with%20enhanced%20intracellular%20hepatitis%20C%20virus%20production%20and%20evasion%20of%20virus%20related-cell%20cycle%20arrest&rft.jtitle=PloS%20one&rft.au=Murayama,%20Asako&rft.date=2012-12-20&rft.volume=7&rft.issue=12&rft.spage=e52697&rft.epage=e52697&rft.pages=e52697-e52697&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0052697&rft_dat=%3Cgale_plos_%3EA477058559%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1327199840&rft_id=info:pmid/23285155&rft_galeid=A477058559&rft_doaj_id=oai_doaj_org_article_968475d952d14e68a9db156dba57fca4&rfr_iscdi=true |