CRISPR/Cas9 介导的里氏木霉基因组编辑。

CRISPR/Cas9-Mediated Genome Editing of Trichoderma reesei.

机构信息

CAS-Key Laboratory of Synthetic Biology, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.

出版信息

Methods Mol Biol. 2021;2234:87-98. doi: 10.1007/978-1-0716-1048-0_8.

DOI:10.1007/978-1-0716-1048-0_8

PMID:33165782

Abstract

In this protocol, we describe the establishment of a CRISPR/Cas9 system in Trichoderma reesei by generating a specific, codon-optimized Cas9-expressing strain and by in vitro transcription of a gRNA. This system induces mutagenesis or introduces a gene in a targeted way based on PEG-mediated protoplast transformation. Up to three targets, multiplexed genome editing can be obtained in one transformation.

摘要

在本方案中，我们通过生成特定的、密码子优化的 Cas9 表达菌株和体外转录 gRNA，在里氏木霉中建立了 CRISPR/Cas9 系统。该系统基于 PEG 介导的原生质体转化，以诱导突变或有针对性地引入基因。在一次转化中，最多可以获得三个靶标，实现多重基因组编辑。