Journal
ELIFE
Volume 9, Issue -, Pages -Publisher
ELIFE SCIENCES PUBLICATIONS LTD
DOI: 10.7554/eLife.56008
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Funding
- Chinese National Program on Key Basic Research Project [2018YFC2000100, 2017YFA0103300, 2016YFA0100400]
- National Science Foundation of China [31871438, 81972457, 31721003, 31871446]
- Fundamental Research Funds for the Central Universities
- Program of Shanghai Academic Research Leader [19XD1403000]
- Shuguang Program of Shanghai Education Development Foundation [19SG18]
- Shanghai Municipal Education Commission [19SG18]
- key project of the Science and Technology of Shanghai Municipality [19JC1415300]
- Shanghai Rising-Star Program [19QA1409600]
- Shanghai Municipal Medical and Health Discipline Construction Projects [2017ZZ02015]
- Young Elite Scientist Sponsorship Program by CAST [2018QNRC001]
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Directly modulating the choice between homologous recombination (HR) and non-homologous end joining (NHEJ) - two independent pathways for repairing DNA double-strand breaks (DSBs) - has the potential to improve the efficiency of gene targeting by CRISPR/Cas9. Here, we have developed a rapid and easy-to-score screening approach for identifying small molecules that affect the choice between the two DSB repair pathways. Using this tool, we identified a small molecule, farrerol, that promotes HR but does not affect NHEJ. Further mechanistic studies indicate that farrerol functions through stimulating the recruitment of RAD51 to DSB sites. Importantly, we demonstrated that farrerol effectively promotes precise targeted integration in human cells, mouse cells and mouse embryos at multiple genomic loci. In addition, treating cells with farrerol did not have any obvious negative effect on genomic stability. Moreover, farrerol significantly improved the knock-in efficiency in blastocysts, and the subsequently generated knock-in mice retained the capacity for germline transmission.
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