数字农科院2.0

Seeing Red, Selecting True: RUBY-Reported Seed Marker Streamlines CRISPR-Clean Rice Breeding

文献类型： 外文期刊

作者： Jin Lei Liu;Tao Yang;Yu Wei Fu;Zhitian Zhan;Hong Chen;Han Cheng;Jiankun Zhou;Yi Ming Wang;Li Jun Tang;Wen Qiang Chen;Ming Wei Wu;Dake Zhao;Chun Ming Liu;Yubing He;Jinxin Liu

作者机构：

关键词： CRISPR-Cas9;Genome editing;RUBY;Transgene-free;Visible marker-assisted selection

期刊名称： Rice

ISSN： 1939-8425

年卷期： 2025 年 18 卷 1 期

页码：

收录情况： SCIE（2025版）

摘要： The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR-associated protein (Cas)9 genome-editing technology has become a cornerstone for generating knockout mutations in plant functional genomics. To obtain genetically stable CRISPR-edited plants, the removal of exogenous CRISPR constructs through genetic segregation is imperative. However, current transgene-free strategies lack universality and operational simplicity. Here, we developed a modular CRISPR toolkit integrated with the widely applicable visual RUBY marker. This system achieved 100% editing efficiency in three independent CRISPR-Cas9 editing events in rice (cv. Zhonghua 11), enabled rapid visual identification of transgene-free progeny, and may provide a framework for future adaptation of CRISPR vectors to other plant species. Our design significantly accelerates the identification of edited lines while bypassing laborious molecular validation steps.

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