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Updated: Jul 16, 2025

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基于CRISPR/Cas9的Nicotiana tabacum基因向突变体的高吞吐量创造

Yudong Chen¹, Haiying Xiang¹, Ling Jia²

¹Yunnan Academy of Tobacco Science, Kunming, 650106, China.

Plant cell reports

|September 12, 2023

概括

研究人员使用4,382个sgRNA为1,060个基因生成了超过10,000个向烟草突变. 优化实验解决了遗传转换的挑战,推进了植物基因编辑能力.

科学领域:

植物遗传学植物遗传学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

烟草 (Nicotiana tabacum) 是植物科学中一个重要的模型生物.
有效的基因操纵对于理解基因功能和作物改进至关重要.

研究的目的:

建立一个高通量系统,用于在烟草中产生向基因突变.
优化基因转换协议以提高效率.

主要方法:

4,382个单导向RNAs (sgRNAs) 的设计和合成,针对1,060个不同的烟草基因.
应用高通量方法来创建10682个向突变系.
进行四个不同的优化实验以解决遗传转换瓶.

主要成果:

成功获得了4382个sgRNAs的综合收集.
创建了一个大规模的 10,682 个目标烟草突变的图书馆.
通过优化实验,确定了提高基因转换效率的关键因素.

结论:

该研究提供了一个强大的高通量平台,用于烟草中大规模的向突变发生.
开发的优化策略显著提高了这一物种遗传转变的成功率.
该资源加速了功能性基因组研究和烟草和潜在的其他作物植物的育种工作.

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CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...