Genome editing by miniature CRISPR/Cas12f1 enzyme in Escherichia coli
Genome editing by miniature CRISPR/Cas12f1 enzyme in Escherichia coli
复制标题
DOI：
10.1016/j.jbiosc.2021.04.009
复制
发表时间：
2021-07-22
影响因子：
2.8
通讯作者：
Honda, Kohsuke
中科院分区：
文献类型：
--
作者：
Okano, Kenji;Sato, Yu;Honda, Kohsuke
成簇规则间隔短回文重复序列（CRISPR）/CRISPR相关（Cas）系统是微生物的有价值的基因组编辑工具。然而，常用的来源于化脓性链球菌的Cas9核酸酶（SpCas 9）由于其细胞毒性和大尺寸（1368个氨基酸[aa]）而不适用于许多工业相关细菌。我们开发了一种替代的基因组编辑系统，使用来自未培养古菌Un 1Cas 12 f1的微型Cas 12 f1核酸酶（529 aa）。当在大肠杆菌MG 1655和BW 25113中编辑四个pcr基因时，CRISPR/Un 1Cas 12 f1系统显示出比CRISPR/SpCas 9系统（50%- 79%）更高的效率（63%-100%）。CRISPR/Un 1Cas 12 f1基因组编辑系统有望应用于多种细菌的基因组编辑。（C）2021，生物技术学会，日本。All rights reserved.
The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system is a valuable genome editing tool for microorganisms. However, the commonly used Cas9 nuclease derived from Streptococcus pyogenes (SpCas9) is not applicable to many industrially relevant bacteria, due to its cytotoxicity and large size (1368 amino acids [aa]). We developed an alternative genome editing system using a miniature Cas12f1 nuclease (529 aa) derived from an uncultured archaeon, Un1Cas12f1. When editing four dispensable genes in Escherichia coli MG1655 and BW25113, the CRISPR/Un1Cas12f1 system showed higher efficiency (63%-100%) than the CRISPR/SpCas9 system (50%- 79%). The CRISPR/Un1Cas12f1 genome editing system is expected to be applied to the genome editing of a wide variety of bacteria. (C ) 2021, The Society for Biotechnology, Japan. All rights reserved.