School of Life Science and Technology, ShanghaiTech University, Shanghai, China.
Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China.
Nat Biotechnol. 2018 Apr;36(4):324-327. doi: 10.1038/nbt.4102. Epub 2018 Mar 19.
The targeting range of CRISPR-Cas9 base editors (BEs) is limited by their G/C-rich protospacer-adjacent motif (PAM) sequences. To overcome this limitation, we developed a CRISPR-Cpf1-based BE by fusing the rat cytosine deaminase APOBEC1 to a catalytically inactive version of Lachnospiraceae bacterium Cpf1. The base editor recognizes a T-rich PAM sequence and catalyzes C-to-T conversion in human cells, while inducing low levels of indels, non-C-to-T substitutions and off-target editing.
CRISPR-Cas9 碱基编辑器 (BE) 的靶向范围受到其富含 G/C 的原间隔基序 (PAM) 序列的限制。为了克服这一限制，我们通过将大鼠胞嘧啶脱氨酶 APOBEC1 融合到无活性的 Lachnospiraceae 细菌 Cpf1 上，开发了一种基于 CRISPR-Cpf1 的 BE。该碱基编辑器识别富含 T 的 PAM 序列，并在人细胞中催化 C 到 T 的转换，同时诱导低水平的插入缺失、非 C 到 T 的取代和脱靶编辑。
