Immortalization of primary marmoset skin fibroblasts by CRISPR-Cas9-mediated gene targeting
ABSTRACTImmortalized cell lines can be used for diverse in vitro experiments, providing invaluable data before conducting in vivo studies Callithrix jacchus, the common marmoset, is a non-human primate model utilized for studying various human diseases. However, only a few immortalized marmoset cell...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2022-11-01
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| Series: | Animal Cells and Systems |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/19768354.2022.2151509 |
| _version_ | 1827937270717480960 |
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| author | Yeon-Ju Jeong Jeongin Cho Jina Kwak Young Hoon Sung Byeong-Cheol Kang |
| author_facet | Yeon-Ju Jeong Jeongin Cho Jina Kwak Young Hoon Sung Byeong-Cheol Kang |
| author_sort | Yeon-Ju Jeong |
| collection | DOAJ |
| description | ABSTRACTImmortalized cell lines can be used for diverse in vitro experiments, providing invaluable data before conducting in vivo studies Callithrix jacchus, the common marmoset, is a non-human primate model utilized for studying various human diseases. However, only a few immortalized marmoset cell lines are currently available. In the present study, we reveal that CRISPR-Cas9-mediated targeting of the p53 gene or CDKN2A locus is an effective means for immortalizing primary marmoset skin fibroblasts. In addition to frameshift mutations that result in premature stop codons, in-frame mutations potentially destroying the DNA-binding motif of p53 are frequently detected in immortalized cells. Like Cdkn2a-deficient mouse cells, CDKN2A-deficient marmoset cells express wild-type p53 proteins normally respond to genotoxic stresses, including adriamycin and etoposide. Taken together, these findings indicate that Cas9- mediated gene targeting of the p53 gene or CDKN2A locus is an effective tool for establishing immortalized marmoset cell lines with defined genetic alterations. |
| first_indexed | 2024-03-13T08:23:06Z |
| format | Article |
| id | doaj.art-ca6b5b60783f4ef8ac16a091f5d3bca6 |
| institution | Directory Open Access Journal |
| issn | 1976-8354 2151-2485 |
| language | English |
| last_indexed | 2024-03-13T08:23:06Z |
| publishDate | 2022-11-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Animal Cells and Systems |
| spelling | doaj.art-ca6b5b60783f4ef8ac16a091f5d3bca62023-05-31T05:33:12ZengTaylor & Francis GroupAnimal Cells and Systems1976-83542151-24852022-11-0126626627410.1080/19768354.2022.2151509Immortalization of primary marmoset skin fibroblasts by CRISPR-Cas9-mediated gene targetingYeon-Ju Jeong0Jeongin Cho1Jina Kwak2Young Hoon Sung3Byeong-Cheol Kang4Convergence Medicine Research Center, Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of KoreaConvergence Medicine Research Center, Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of KoreaGraduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of KoreaConvergence Medicine Research Center, Asan Institute for Life Sciences, Asan Medical Center, Seoul, Republic of KoreaGraduate School of Translational Medicine, Seoul National University College of Medicine, Seoul, Republic of KoreaABSTRACTImmortalized cell lines can be used for diverse in vitro experiments, providing invaluable data before conducting in vivo studies Callithrix jacchus, the common marmoset, is a non-human primate model utilized for studying various human diseases. However, only a few immortalized marmoset cell lines are currently available. In the present study, we reveal that CRISPR-Cas9-mediated targeting of the p53 gene or CDKN2A locus is an effective means for immortalizing primary marmoset skin fibroblasts. In addition to frameshift mutations that result in premature stop codons, in-frame mutations potentially destroying the DNA-binding motif of p53 are frequently detected in immortalized cells. Like Cdkn2a-deficient mouse cells, CDKN2A-deficient marmoset cells express wild-type p53 proteins normally respond to genotoxic stresses, including adriamycin and etoposide. Taken together, these findings indicate that Cas9- mediated gene targeting of the p53 gene or CDKN2A locus is an effective tool for establishing immortalized marmoset cell lines with defined genetic alterations.https://www.tandfonline.com/doi/10.1080/19768354.2022.2151509Marmosetimmortalizationcell lineCas9genotoxic stress |
| spellingShingle | Yeon-Ju Jeong Jeongin Cho Jina Kwak Young Hoon Sung Byeong-Cheol Kang Immortalization of primary marmoset skin fibroblasts by CRISPR-Cas9-mediated gene targeting Animal Cells and Systems Marmoset immortalization cell line Cas9 genotoxic stress |
| title | Immortalization of primary marmoset skin fibroblasts by CRISPR-Cas9-mediated gene targeting |
| title_full | Immortalization of primary marmoset skin fibroblasts by CRISPR-Cas9-mediated gene targeting |
| title_fullStr | Immortalization of primary marmoset skin fibroblasts by CRISPR-Cas9-mediated gene targeting |
| title_full_unstemmed | Immortalization of primary marmoset skin fibroblasts by CRISPR-Cas9-mediated gene targeting |
| title_short | Immortalization of primary marmoset skin fibroblasts by CRISPR-Cas9-mediated gene targeting |
| title_sort | immortalization of primary marmoset skin fibroblasts by crispr cas9 mediated gene targeting |
| topic | Marmoset immortalization cell line Cas9 genotoxic stress |
| url | https://www.tandfonline.com/doi/10.1080/19768354.2022.2151509 |
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