HDAC inhibitors improve CRISPR-mediated HDR editing efficiency in iPSCs

被引：20

作者：

Zhang, Jian-Ping ^{[1
]}

Yang, Zhi-Xue ^{[1
]}

Zhang, Feng ^{[1
]}

Fu, Ya-Wen ^{[1
]}

Dai, Xin-Yue ^{[1
]}

Wen, Wei ^{[1
]}

Zhang, Beldon ^{[2
]}

Choi, Hannah ^{[3
]}

Chen, Wanqiu ^{[3
]}

Brown, Meredith ^{[3
]}

Baylink, David ^{[2
]}

Zhang, Lei ^{[1
,4
,7
]}

Qiu, Hongyu ^{[8
]}

Wang, Charles ^{[3
]}

Cheng, Tao ^{[1
,5
,6
]}

Zhang, Xiao-Bing ^{[1
,2
]}

机构：

[1] Chinese Acad Med Sci & Peking Union Med Coll, Natl Clin Res Ctr Blood Dis, Inst Hematol & Blood Dis Hosp, State Key Lab Expt Hematol, Tianjin 300020, Peoples R China

[2] Loma Linda Univ, Sch Med, Loma Linda, CA 92354 USA

[3] Loma Linda Univ, Sch Med, Ctr Genom, Loma Linda, CA 92350 USA

[4] CAMS Key Lab Gene Therapy Blood Dis, Tianjin 300020, Peoples R China

[5] Chinese Acad Med Sci, Ctr Stem Cell Med, Tianjin 300020, Peoples R China

[6] Peking Union Med Coll, Dept Stem Cell & Regenerat Med, Tianjin 300020, Peoples R China

[7] Tianjin Key Lab Gene Therapy Blood Dis, Tianjin 300020, Peoples R China

[8] Georgia State Univ, Inst Biomed Sci, Ctr Mol & Translat Med, Atlanta, GA 30303 USA

来源：

SCIENCE CHINA-LIFE SCIENCES | 2021年 / 64卷 / 09期

基金：

中国国家自然科学基金;

关键词：

HDAC inhibitors; CRISPR-Cas9; genome editing; iPSC; PLURIPOTENT STEM-CELLS; HISTONE DEACETYLASE INHIBITOR; CHROMATIN; GENE; ACTIVATION; CHALLENGES; DISEASE;

D O I：

10.1007/s11427-020-1855-4

中图分类号：

Q [生物科学];

学科分类号：

07 ; 0710 ; 09 ;

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 (similar to 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. 1.1-fold change). These studies provide a new strategy for HDR-editing of silent genes in iPSCs.

引用

页码：1449 / 1462

页数：14

共 56 条

[1] The clinical potential of gene editing as a tool to engineer cell-based therapeutics [J].