CRISPR/LbCas12a-Mediated Genome Editing in Soybean.

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2023-01-01 DOI:10.1007/978-1-0716-3131-7_3

Dawei Liang, Yubo Liu, Chao Li, Qin Wen, Jianping Xu, Lizhao Geng, Chunxia Liu, Huaibing Jin, Yang Gao, Heng Zhong, John Dawson, Bin Tian, Brenden Barco, Xiujuan Su, Shujie Dong, Changbao Li, Sivamani Elumalai, Qiudeng Que, Ian Jepson, Liang Shi

引用次数: 2

Abstract

Currently methods for generating soybean edited lines are time-consuming, inefficient, and limited to certain genotypes. Here we describe a fast and highly efficient genome editing method based on CRISPR-Cas12a nuclease system in soybean. The method uses Agrobacterium-mediated transformation to deliver editing constructs and uses aadA or ALS genes as selectable marker. It only takes about 45 days to obtain greenhouse-ready edited plants at higher than 30% transformation efficiency and 50% editing rate. The method is applicable to other selectable markers including EPSPS and has low transgene chimera rate. The method is also genotype-flexible and has been applied to genome editing of several elite soybean varieties.

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CRISPR/LbCas12a 介导的大豆基因组编辑。

目前生成大豆编辑品系的方法耗时长、效率低，而且仅限于某些基因型。在此，我们介绍了一种基于 CRISPR-Cas12a 核酸酶系统的大豆快速高效基因组编辑方法。该方法利用农杆菌介导的转化来传递编辑构建体，并使用 aadA 或 ALS 基因作为可选择标记。只需约 45 天即可获得温室就绪的编辑植株，转化效率高于 30%，编辑率高于 50%。该方法适用于包括 EPSPS 在内的其他可选择标记，且转基因嵌合率低。该方法还具有基因型灵活性，已被应用于多个优良大豆品种的基因组编辑。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.