Assaying break and nick-induced homologous recombination in mammalian cells using the DR-GFP reporter and cas9 nucleases Journal Article
| Authors: | Vriend, L. E. M.; Jasin, M.; Krawczyk, P. M. |
| Article Title: | Assaying break and nick-induced homologous recombination in mammalian cells using the DR-GFP reporter and cas9 nucleases |
| Abstract: | Thousands of DNA breaks occur daily in mammalian cells, including potentially tumorigenic double-strand breaks (DSBs) and less dangerous but vastly more abundant single-strand breaks (SSBs). The majority of SSBs are quickly repaired, but some can be converted to DSBs, posing a threat to the integrity of the genome. Although SSBs are usually repaired by dedicated pathways, they can also trigger homologous recombination (HR), an error-free pathway generally associated with DSB repair. While HR-mediated DSB repair has been extensively studied, the mechanisms of HR-mediated SSB repair are less clear. This chapter describes a protocol to investigate SSB-induced HR in mammalian cells employing the DR-GFP reporter, which has been widely used in DSB repair studies, together with an adapted bacterial CRISPR/Cas system. © 2014 Elsevier Inc. All rights reserved. |
| Keywords: | mammalia; homologous recombination; dna repair; bacteria (microorganisms); crispr/cas; cas9; dna nicks; dr-gfp reporter; single-strand breaks |
| Journal Title: | Methods in Enzymology |
| Volume: | 546 |
| ISSN: | 0076-6879 |
| Publisher: | Academic Press |
| Date Published: | 2014-01-01 |
| Start Page: | 175 |
| End Page: | 191 |
| Language: | English |
| DOI: | 10.1016/b978-0-12-801185-0.00009-x |
| PROVIDER: | scopus |
| PUBMED: | 25398341 |
| PMCID: | PMC4408992 |
| DOI/URL: | |
| Notes: | Chapter 9 in "The Use of CRISPR/Cas9, ZFNs, and TALENs in Generating Site-Specific Genome Alterations" (ISBN: 978-0-12-801185-0) Export Date: 2 April 2015 -- Source: Scopus |
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