Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair
Searching for factors to improve knockin efficiency for therapeutic applications, biotechnology, and generation of non-human primate models of disease, we found that the strand exchange protein RAD51 can significantly increase Cas9-mediated homozygous knockin in mouse embryos through an interhomolog...
| Main Authors: | , , , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
Elsevier BV
2022
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| Online Access: | https://hdl.handle.net/1721.1/143451 |
| _version_ | 1826216819215564800 |
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| author | Wilde, Jonathan J Aida, Tomomi del Rosario, Ricardo CH Kaiser, Tobias Qi, Peimin Wienisch, Martin Zhang, Qiangge Colvin, Steven Feng, Guoping |
| author2 | Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences |
| author_facet | Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Wilde, Jonathan J Aida, Tomomi del Rosario, Ricardo CH Kaiser, Tobias Qi, Peimin Wienisch, Martin Zhang, Qiangge Colvin, Steven Feng, Guoping |
| author_sort | Wilde, Jonathan J |
| collection | MIT |
| description | Searching for factors to improve knockin efficiency for therapeutic applications, biotechnology, and generation of non-human primate models of disease, we found that the strand exchange protein RAD51 can significantly increase Cas9-mediated homozygous knockin in mouse embryos through an interhomolog repair (IHR) mechanism. IHR is a hallmark of meiosis but only occurs at low frequencies in somatic cells, and its occurrence in zygotes is controversial. Using multiple approaches, we provide evidence for an endogenous IHR mechanism in the early embryo that can be enhanced by RAD51. This process can be harnessed to generate homozygotes from wild-type zygotes using exogenous donors and to convert heterozygous alleles into homozygous alleles without exogenous templates. Furthermore, we identify additional IHR-promoting factors and describe features of IHR events. Together, our findings show conclusive evidence for IHR in mouse embryos and describe an efficient method for enhanced gene conversion. |
| first_indexed | 2024-09-23T16:53:36Z |
| format | Article |
| id | mit-1721.1/143451 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T16:53:36Z |
| publishDate | 2022 |
| publisher | Elsevier BV |
| record_format | dspace |
| spelling | mit-1721.1/1434512023-01-11T20:43:09Z Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair Wilde, Jonathan J Aida, Tomomi del Rosario, Ricardo CH Kaiser, Tobias Qi, Peimin Wienisch, Martin Zhang, Qiangge Colvin, Steven Feng, Guoping Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences McGovern Institute for Brain Research at MIT Searching for factors to improve knockin efficiency for therapeutic applications, biotechnology, and generation of non-human primate models of disease, we found that the strand exchange protein RAD51 can significantly increase Cas9-mediated homozygous knockin in mouse embryos through an interhomolog repair (IHR) mechanism. IHR is a hallmark of meiosis but only occurs at low frequencies in somatic cells, and its occurrence in zygotes is controversial. Using multiple approaches, we provide evidence for an endogenous IHR mechanism in the early embryo that can be enhanced by RAD51. This process can be harnessed to generate homozygotes from wild-type zygotes using exogenous donors and to convert heterozygous alleles into homozygous alleles without exogenous templates. Furthermore, we identify additional IHR-promoting factors and describe features of IHR events. Together, our findings show conclusive evidence for IHR in mouse embryos and describe an efficient method for enhanced gene conversion. 2022-06-15T17:22:38Z 2022-06-15T17:22:38Z 2021 2022-06-15T17:20:15Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/143451 Wilde, Jonathan J, Aida, Tomomi, del Rosario, Ricardo CH, Kaiser, Tobias, Qi, Peimin et al. 2021. "Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair." Cell, 184 (12). en 10.1016/J.CELL.2021.04.035 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier BV PMC |
| spellingShingle | Wilde, Jonathan J Aida, Tomomi del Rosario, Ricardo CH Kaiser, Tobias Qi, Peimin Wienisch, Martin Zhang, Qiangge Colvin, Steven Feng, Guoping Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair |
| title | Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair |
| title_full | Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair |
| title_fullStr | Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair |
| title_full_unstemmed | Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair |
| title_short | Efficient embryonic homozygous gene conversion via RAD51-enhanced interhomolog repair |
| title_sort | efficient embryonic homozygous gene conversion via rad51 enhanced interhomolog repair |
| url | https://hdl.handle.net/1721.1/143451 |
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