文献类型：期刊文献

英文题名：A simple approach to mediate genome editing in the filamentous fungus Trichoderma reesei by CRISPR/Cas9-coupled in vivo gRNA transcription

作者：Wu, Chuan[1];Chen, Yumeng[1];Qiu, Yifei[1];Niu, Xiao[1];Zhu, Ningjian[2];Chen, Jiehui[1];Yao, Hong[1];Wang, Wei[1];Ma, Yushu[1]

机构：[1]East China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd,POB 311, Shanghai 200237, Peoples R China;[2]East China Normal Univ, Affiliated Middle Sch 1, Shanghai 200086, Peoples R China

年份：2020

卷号：42

期号：7

起止页码：1203

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;EI(收录号：20201708512120);WOS:【SCI-EXPANDED(收录号:WOS:000538373400011)】；

语种：英文

外文关键词：Filamentous fungus; Trichoderma reesei; CRISPR; Cas9; In vivo gRNA transcription; Gene editing

摘要：Objective To simplify CRISPR/Cas9 genome editing in the industrial filamentous fungus Trichoderma reesei based on in vivo guide RNA (gRNA) transcription. Results Two putative RNA polymerase III U6 snRNA genes were identified in the genome of T. reesei QM6a by BLASTN using Myceliophthora. thermophila U6 snRNA gene as the template. The regions approximately 500 bp upstream of two U6 genes were efficient promoters for the in vivo expression of gRNA. The CRISPR system consisting of Cas9 and in vivo synthesized gRNA under control of the T. reesei U6 snRNA promoters was sufficient to cause a frameshift mutation in the ura5 gene via non-homologous end-joining-mediated events. Conclusions We report a simple gene editing method using a CRISPR/Cas9-coupled in vivo gRNA transcription system in T. reesei.