Analysis of the Effectiveness of CRISPR/Cas9-Editing of the GEX2 Gene by Ribonucleoprotein Complexes in Maize Protoplasts

IF 0.6 4区 生物学 Q4 GENETICS & HEREDITY
E. M. Moiseeva, V. V. Fadeev, Yu. V. Fadeeva, Yu. S. Gusev, M. I. Chumakov
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引用次数: 0

Abstract

The GEX2 protein is expressed in the maize gamete membranes and is necessary during fertilization at the stage of contact (adhesion) of gamete membranes. Knockout of GEX2 gene can presumably lead to impaired fertilization and, as a result, to the formation of matroclinic haploid maize embryos. The purpose of this study is to analyze the efficiency of CRISPR/Cas9 editing of the GEX2 gene after PEG-mediated transfection of maize protoplasts by ribonucleoprotein (RNP) complexes with different gRNAs. For the first time, constructs for CRISPR/Cas9 editing of the maize GEX2 gene have been created, the effectiveness of which has been proven on protoplasts and reaches 10.7%, depending on the selected gRNA and the ratio and number of components in the RNP complexes.

Abstract Image

玉米原生质体中核糖核蛋白复合物对 GEX2 基因的 CRISPR/Cas9 编辑效果分析
摘要-GEX2 蛋白在玉米配子膜中表达,是受精过程中配子膜接触(粘连)阶段所必需的。敲除 GEX2 基因可能会导致受精障碍,从而形成母系单倍体玉米胚胎。本研究的目的是分析不同 gRNA 的核糖核蛋白(RNP)复合物通过 PEG 介导转染玉米原生质体后 CRISPR/Cas9 编辑 GEX2 基因的效率。我们首次创建了对玉米GEX2基因进行CRISPR/Cas9编辑的构建体,其有效性已在原生质体上得到证实,达到10.7%,这取决于所选的gRNA以及RNP复合物中成分的比例和数量。
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来源期刊
Russian Journal of Genetics
Russian Journal of Genetics 生物-遗传学
CiteScore
1.00
自引率
33.30%
发文量
126
审稿时长
1 months
期刊介绍: Russian Journal of Genetics is a journal intended to make significant contribution to the development of genetics. The journal publishes reviews and experimental papers in the areas of theoretical and applied genetics. It presents fundamental research on genetic processes at molecular, cell, organism, and population levels, including problems of the conservation and rational management of genetic resources and the functional genomics, evolutionary genomics and medical genetics.
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