Journal
GENOME BIOLOGY
Volume 20, Issue -, Pages -Publisher
BMC
DOI: 10.1186/s13059-019-1762-8
Keywords
CRISPR; Cas9; Chromatin accessibility; Transcription activation domain; Proximal dsgRNA
Funding
- National Transgenic Science and Technology Program of China [2016ZX08010-002, 2018ZX0801002B]
- National Key Research and Development Program of China [2016YFD0100602]
- Chinese Academy of Sciences [XDB11030500]
- National Natural Science Foundation of China [31700314, 31672015]
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The CRISPR/Cas9 system is unable to edit all targetable genomic sites with full efficiency in vivo. We show that Cas9-mediated editing is more efficient in open chromatin regions than in closed chromatin regions in rice. A construct (Cas9-TV) formed by fusing a synthetic transcription activation domain to Cas9 edits target sites more efficiently, even in closed chromatin regions. Moreover, combining Cas9-TV with a proximally binding dead sgRNA (dsgRNA) further improves editing efficiency up to several folds. The use of Cas9-TV/dsgRNA thus provides a novel strategy for obtaining efficient genome editing in vivo, especially at nuclease-refractory target sites.
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