Modulating chromatin accessibility by transactivation and targeting proximal dsgRNAs enhances Cas9 editing efficiency in vivo
Modulating chromatin accessibility by transactivation and targeting proximal dsgRNAs enhances Cas9 editing efficiency in vivo
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通过反式激活和靶向近端 dsgRNA 调节染色质可及性可增强 Cas9 体内编辑效率
DOI：
10.1186/s13059-019-1762-8
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发表时间：
2019-07-26
期刊：
影响因子：
12.3
通讯作者：
Qiu, Jin-Long
中科院分区：
文献类型：
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作者：
Liu, Guanwen;Yin, Kangquan;Qiu, Jin-Long
CRISPR/CAS9系统无法在体内具有完全效率的所有目标基因组站点编辑。我们表明，在开放式染色质区域中，CAS9介导的编辑比大米中的封闭染色质区域更有效。即使在封闭的染色质区域中，将合成转录激活域融合到CAS9编辑目标位点的构建体（CAS9-TV）也会更有效地编辑目标位点。此外，将CAS9-TV与近端结合的死sgrna（DSGRNA）相结合，进一步提高了编辑效率，最高为几倍。因此，CAS9-TV/DSGRNA的使用提供了一种新的策略，用于在体内获得有效的基因组编辑，尤其是在核酸酶 - 饮食靶点位点。
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.