HDAC inhibitors improve CRISPR-mediated HDR editing efficiency in iPSCs  

作　　者：Jian-Ping Zhang Zhi-Xue Yang Feng Zhang Ya-Wen Fu Xin-Yue Dai Wei Wen Beldon Zhang Hannah Choi Wanqiu Chen Meredith Brown David Baylink Lei Zhang Hongyu Qiu Charles Wang Tao Cheng Xiao-Bing Zhang 

机构地区：[1]State Key Laboratory of Experimental Hematology,National Clinical Research Center for Blood Diseases,Institute of Hematology&Blood Diseases Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Tianjin 300020,China [2]School of Medicine,Loma Linda University,Loma Linda,CA 92354,USA [3]Center for Genomics,School of Medicine,Loma Linda University,Loma Linda,CA 92350,USA [4]CAMS Key Laboratory of Gene Therapy for Blood Diseases,Tianjin 300020,China [5]Center for Stem Cell Medicine,Chinese Academy of Medical Sciences,Tianjin 300020,China [6]Department of Stem Cell&Regenerative Medicine,Peking Union Medical College,Tianjin 300020,China [7]Tianjin Key Laboratory of Gene Therapy for Blood Diseases,Tianjin 300020,China [8]Center of Molecular and Translational Medicine,Institution of Biomedical Science,Georgia State University,Atlanta,GA 30303,USA

出　　处：《Science China(Life Sciences)》2021年第9期1449-1462,共14页中国科学（生命科学英文版）

基　　金：supported by the National Natural Science Foundation of China(81870149,82070115,81770198,81700184,81570164,81861148029,81700183,81421002,81890990,81730006);National Key Research and Development Program of China(2019YFA0110803,2019YFA0110204,2016YFA0100600,2017YFA0103400);CAMS Innovation Fund for Medical Sciences(CIFMS)(2017-I2M-B&R-04,2019-I2M-1-006,2017-I2M-1-015,2016-I2M-1-017,2017-I2M-2-001);Ministry of Science and Technology of China(2015CB964902,2015CB964400);Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences(2018PT31004);CAMS Key Laboratory of Gene Therapy for Blood Diseases(2017PT31047,2018PT31038);American Heart Association(18IPA34170301)。

摘　　要：Genome-edited human induced pluripotent stem cells(iPSCs)hold great promise for therapeutic applications.However,low editing efficiency has hampered the applications of CRISPR-Cas9 technology in creating knockout and homology-directed repair(HDR)-edited iPSC lines,particularly for silent genes.This is partially due to chromatin compaction,inevitably limiting Cas9 access to the target DNA.Among the six HDAC inhibitors we examined,vorinostat,or suberoylanilide hydroxamic acid(SAHA),led to the highest HDR efficiency at both open and closed loci,with acceptable toxicity.HDAC inhibitors equally increased non-homologous end joining(NHEJ)editing efficiencies(~50%)at both open and closed loci,due to the considerable HDAC inhibitor-mediated increase in Cas9 and sgRNA expression.However,we observed more substantial HDR efficiency improvement at closed loci relative to open chromatin(2.8 vs.1.7-fold change).These studies provide a new strategy for HDRediting of silent genes in iPSCs.

关 键 词：HDAC inhibitors CRISPR-Cas9 genome editing IPSC 

分 类 号：Q78[生物学—分子生物学]