一种具有广泛 PAM 范围、高特异性和高活性的工程化 ScCas9。
An engineered ScCas9 with broad PAM range and high specificity and activity.
机构信息
MIT Center for Bits and Atoms, Massachusetts Institute of Technology, Cambridge, MA, USA.
MIT Media Lab, Massachusetts Institute of Technology, Cambridge, MA, USA.
出版信息
Nat Biotechnol. 2020 Oct;38(10):1154-1158. doi: 10.1038/s41587-020-0517-0. Epub 2020 May 11.
CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. In the present study, we combine protein motifs from several orthologs to engineer two variants of Streptococcus canis Cas9-Sc and a higher-fidelity mutant HiFi-Sc-that have simultaneously broad 5'-NNG-3' PAM compatibility, robust DNA-cleavage activity and minimal off-target activity. Sc and HiFi-Sc extend the use of CRISPR editing for diverse applications.
CRISPR 酶在靶位切割位点附近需要一个前导间隔相邻基序（PAM），这限制了可编辑的序列。在本研究中，我们将来自几个同源物的蛋白质基序进行组合，工程改造了两种链球菌 Cas9-Sc 变体和一种具有更高保真度的突变体 HiFi-Sc，它们同时具有广泛的 5'-NNG-3' PAM 兼容性、强大的 DNA 切割活性和最小的脱靶活性。Sc 和 HiFi-Sc 扩展了 CRISPR 编辑的用途，适用于各种应用。
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