TY - JOUR TI - A high-throughput small molecule screen identifies farrerol as a potentiator of CRISPR/Cas9-mediated genome editing AU - Zhang, Weina AU - Chen, Yu AU - Yang, Jiaqing AU - Zhang, Jing AU - Yu, Jiayu AU - Wang, Mengting AU - Zhao, Xiaodong AU - Wei, Ke AU - Wan, Xiaoping AU - Xu, Xiaojun AU - Jiang, Ying AU - Chen, Jiayu AU - Gao, Shaorong AU - Mao, Zhiyong A2 - Heyer, Wolf-Dietrich A2 - Tyler, Jessica K A2 - Heyer, Wolf-Dietrich VL - 9 PY - 2020 DA - 2020/07/09 SP - e56008 C1 - eLife 2020;9:e56008 DO - 10.7554/eLife.56008 UR - https://doi.org/10.7554/eLife.56008 AB - 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. KW - CRISPR/Cas9 KW - knock-in KW - homologous recombination (HR) KW - non-homologous end joining (NHEJ) KW - farrerol JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -