Interhomolog homologous recombination in mouse embryonic stem cells Journal Article
| Authors: | Vanoli, F.; Prakash, R.; White, T.; Jasin, M. |
| Article Title: | Interhomolog homologous recombination in mouse embryonic stem cells |
| Abstract: | Homologous recombination is a critical mechanism for the repair of DNA double-strand breaks (DSBs). It occurs predominantly between identical sister chromatids and at lower frequency can also occur between homologs. Interhomolog homologous recombination (IH-HR) has the potential lead to substantial loss of genetic information, i.e., loss of heterozygosity (LOH), when it is accompanied by crossing over. In this chapter, we describe a system to study IH-HR induced by a defined DSB in mouse embryonic stem cells derived from F1 hybrid mice. This system is based on the placement of mutant selectable marker genes, one of which contains an I-SceI endonuclease cleavage site, on the two homologs such that repair of the I-SceI-generated DSB from the homolog leads to drug resistance. Loss of heterozygosity arising during IH-HR is analyzed using a PCR-based approach. Finally, we present a strategy to analyze the role of BLM helicase in this system. © 2021, Springer Science+Business Media, LLC, part of Springer Nature. |
| Keywords: | mouse embryonic stem cells; i-scei; homology-directed repair (hdr); bloom helicase (blm); interhomolog homologous recombination (ih-hr); loss of heterozygosity (loh) |
| Journal Title: | Methods in Molecular Biology |
| Volume: | 2153 |
| ISSN: | 1064-3745 |
| Publisher: | Humana Press Inc |
| Date Published: | 2021-01-01 |
| Start Page: | 127 |
| End Page: | 143 |
| Language: | English |
| DOI: | 10.1007/978-1-0716-0644-5_10 |
| PUBMED: | 32840777 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Chapter 10 in "Homologous Recombination: Methods and Protocols" (ISBN: 978-1-0716-0643-8) -- Export Date: 1 October 2020 -- Source: Scopus |
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