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中国科学家研发出新型植物双碱基编辑器
作者:小柯机器人 发布时间:2020/1/16 10:36:49

中国科学院遗传与发育研究所高彩霞团队和李家洋团队合作实现利用胞嘧啶和腺嘌呤双碱基编辑器对植物基因进行定向、随机诱变。该项研究成果2020年1月13日在线发表在《自然—生物技术》上。

研究人员设计了五个饱和的靶向内源诱变编辑器(STEME),它们可以产生从头突变并促进植物基因的定向进化。在水稻原生质体中,STEME-1在同一目标位点编辑胞嘧啶和腺嘌呤,C> T效率高达61.61%,同时C> T和A> G效率高达15.10%。STEMP-NG结合了切口酶Cas9-NG原型间隔子附近的基序变体,与水稻原生质体中的20个单个指导RNA一起使用,可在水稻乙酰基的56个氨基酸部分-辅酶A羧化酶(OsACC)基因产生接近饱和的诱变(73.21%)。研究人员还将STEME-1和STEME-NG用于水稻中OsACC基因的定向进化,并获得了除草剂抗性突变。这套STEME将加快性状的发展,并能够在适合基于CRISPR编辑的任何植物中工作。

据悉,作物基因的定向饱和诱变可用于生产具有改良农艺性能的遗传变异。但是,能够用于植物基因定向进化的工具是有限的,例如易错PCR或DNA移位。

附:英文原文

Title: Targeted, random mutagenesis of plant genes with dual cytosine and adenine base editors

Author: Chao Li, Rui Zhang, Xiangbing Meng, Sha Chen, Yuan Zong, Chunju Lu, Jin-Long Qiu, Yu-Hang Chen, Jiayang Li, Caixia Gao

Issue&Volume: 2020-01-13

Abstract: Targeted saturation mutagenesis of crop genes could be applied to produce genetic variants with improved agronomic performance. However, tools for directed evolution of plant genes, such as error-prone PCR or DNA shuffling, are limited1. We engineered five saturated targeted endogenous mutagenesis editors (STEMEs) that can generate de novo mutations and facilitate directed evolution of plant genes. In rice protoplasts, STEME-1 edited cytosine and adenine at the same target site with C>T efficiency up to 61.61% and simultaneous C>T and A>G efficiency up to 15.10%. STEME-NG, which incorporates the nickase Cas9-NG protospacer-adjacent motif variant, was used with 20 individual single guide RNAs in rice protoplasts to produce near-saturated mutagenesis (73.21%) for a 56-amino-acid portion of the rice acetyl-coenzyme A carboxylase (OsACC). We also applied STEME-1 and STEME-NG for directed evolution of the OsACC gene in rice and obtained herbicide resistance mutations. This set of two STEMEs will accelerate trait development and should work in any plants amenable to CRISPR-based editing.

DOI: 10.1038/s41587-019-0393-7

Source: https://www.nature.com/articles/s41587-019-0393-7

期刊信息

Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:31.864
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex