Journal
SCIENCE CHINA-LIFE SCIENCES
Volume 64, Issue 9, Pages 1449-1462Publisher
SCIENCE PRESS
DOI: 10.1007/s11427-020-1855-4
Keywords
HDAC inhibitors; CRISPR-Cas9; genome editing; iPSC
Categories
Funding
- 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-BR-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]
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Research shows that HDAC inhibitors can increase the efficiency of genome editing, especially when dealing with silent genes. HDR efficiency improvement is more significant at closed loci.
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.
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