Whole-Genome Sequencing of Vero E6 (VERO C1008) and Comparative Analysis of Four Vero Cell Sublines

The Vero cell line is an immortalized cell line established from kidney epithelial cells of the African green monkey. A variety of Vero sublines have been developed and can be classified into four major cell lineages. In this study, we determined the whole-genome sequence of Vero E6 (VERO C1008), wh...

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Main Authors: Kazuhiro Konishi, Toshiyuki Yamaji, Chisato Sakuma, Fumio Kasai, Toshinori Endo, Arihiro Kohara, Kentaro Hanada, Naoki Osada
Format: Article
Language:English
Published: Frontiers Media S.A. 2022-03-01
Series:Frontiers in Genetics
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Online Access:https://www.frontiersin.org/articles/10.3389/fgene.2022.801382/full
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author Kazuhiro Konishi
Toshiyuki Yamaji
Chisato Sakuma
Fumio Kasai
Toshinori Endo
Arihiro Kohara
Kentaro Hanada
Naoki Osada
author_facet Kazuhiro Konishi
Toshiyuki Yamaji
Chisato Sakuma
Fumio Kasai
Toshinori Endo
Arihiro Kohara
Kentaro Hanada
Naoki Osada
author_sort Kazuhiro Konishi
collection DOAJ
description The Vero cell line is an immortalized cell line established from kidney epithelial cells of the African green monkey. A variety of Vero sublines have been developed and can be classified into four major cell lineages. In this study, we determined the whole-genome sequence of Vero E6 (VERO C1008), which is one of the most widely used cell lines for the proliferation and isolation of severe acute respiratory syndrome coronaviruses (SARS-CoVs), and performed comparative analysis among Vero JCRB0111, Vero CCL-81, Vero 76, and Vero E6. Analysis of the copy number changes and loss of heterozygosity revealed that these four sublines share a large deletion and loss of heterozygosity on chromosome 12, which harbors type I interferon and CDKN2 gene clusters. We identified a substantial number of genetic differences among the sublines including single nucleotide variants, indels, and copy number variations. The spectrum of single nucleotide variants indicated a close genetic relationship between Vero JCRB0111 and Vero CCL-81, and between Vero 76 and Vero E6, and a considerable genetic gap between the former two and the latter two lines. In contrast, we confirmed the pattern of genomic integration sites of simian endogenous retroviral sequences, which was consistent among the sublines. We identified subline-specific/enriched loss of function and missense variants, which potentially contribute to the differences in response to viral infection among the Vero sublines. In particular, we identified four genes (IL1RAP, TRIM25, RB1CC1, and ATG2A) that contained missense variants specific or enriched in Vero E6. In addition, we found that V739I variants of ACE2, which functions as the receptor for SARS-CoVs, were heterozygous in Vero JCRB0111, Vero CCL-81, and Vero 76; however, Vero E6 harbored only the allele with isoleucine, resulting from the loss of one of the X chromosomes.
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spelling doaj.art-3f827e65fd024955aec55357bdae0ce52022-12-21T23:54:31ZengFrontiers Media S.A.Frontiers in Genetics1664-80212022-03-011310.3389/fgene.2022.801382801382Whole-Genome Sequencing of Vero E6 (VERO C1008) and Comparative Analysis of Four Vero Cell SublinesKazuhiro Konishi0Toshiyuki Yamaji1Chisato Sakuma2Fumio Kasai3Toshinori Endo4Arihiro Kohara5Kentaro Hanada6Naoki Osada7Graduate School of Information Science and Technology, Hokkaido University, Sapporo, JapanDepartment of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, JapanDepartment of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, JapanLaboratory of Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, JapanFaculty of Information Science and Technology, Hokkaido University, Sapporo, JapanLaboratory of Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Osaka, JapanDepartment of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, JapanFaculty of Information Science and Technology, Hokkaido University, Sapporo, JapanThe Vero cell line is an immortalized cell line established from kidney epithelial cells of the African green monkey. A variety of Vero sublines have been developed and can be classified into four major cell lineages. In this study, we determined the whole-genome sequence of Vero E6 (VERO C1008), which is one of the most widely used cell lines for the proliferation and isolation of severe acute respiratory syndrome coronaviruses (SARS-CoVs), and performed comparative analysis among Vero JCRB0111, Vero CCL-81, Vero 76, and Vero E6. Analysis of the copy number changes and loss of heterozygosity revealed that these four sublines share a large deletion and loss of heterozygosity on chromosome 12, which harbors type I interferon and CDKN2 gene clusters. We identified a substantial number of genetic differences among the sublines including single nucleotide variants, indels, and copy number variations. The spectrum of single nucleotide variants indicated a close genetic relationship between Vero JCRB0111 and Vero CCL-81, and between Vero 76 and Vero E6, and a considerable genetic gap between the former two and the latter two lines. In contrast, we confirmed the pattern of genomic integration sites of simian endogenous retroviral sequences, which was consistent among the sublines. We identified subline-specific/enriched loss of function and missense variants, which potentially contribute to the differences in response to viral infection among the Vero sublines. In particular, we identified four genes (IL1RAP, TRIM25, RB1CC1, and ATG2A) that contained missense variants specific or enriched in Vero E6. In addition, we found that V739I variants of ACE2, which functions as the receptor for SARS-CoVs, were heterozygous in Vero JCRB0111, Vero CCL-81, and Vero 76; however, Vero E6 harbored only the allele with isoleucine, resulting from the loss of one of the X chromosomes.https://www.frontiersin.org/articles/10.3389/fgene.2022.801382/fullVero cellsgenome sequencingVero E6cell linescell substrate
spellingShingle Kazuhiro Konishi
Toshiyuki Yamaji
Chisato Sakuma
Fumio Kasai
Toshinori Endo
Arihiro Kohara
Kentaro Hanada
Naoki Osada
Whole-Genome Sequencing of Vero E6 (VERO C1008) and Comparative Analysis of Four Vero Cell Sublines
Frontiers in Genetics
Vero cells
genome sequencing
Vero E6
cell lines
cell substrate
title Whole-Genome Sequencing of Vero E6 (VERO C1008) and Comparative Analysis of Four Vero Cell Sublines
title_full Whole-Genome Sequencing of Vero E6 (VERO C1008) and Comparative Analysis of Four Vero Cell Sublines
title_fullStr Whole-Genome Sequencing of Vero E6 (VERO C1008) and Comparative Analysis of Four Vero Cell Sublines
title_full_unstemmed Whole-Genome Sequencing of Vero E6 (VERO C1008) and Comparative Analysis of Four Vero Cell Sublines
title_short Whole-Genome Sequencing of Vero E6 (VERO C1008) and Comparative Analysis of Four Vero Cell Sublines
title_sort whole genome sequencing of vero e6 vero c1008 and comparative analysis of four vero cell sublines
topic Vero cells
genome sequencing
Vero E6
cell lines
cell substrate
url https://www.frontiersin.org/articles/10.3389/fgene.2022.801382/full
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