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

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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Publication Details

Journal
Rice
Published
2025-10-03
DOI
https://doi.org/10.1186/s12284-025-00841-0
Citations
2
Primary Topic
CRISPR and Genetic Engineering
Type
article
Field-Weighted Citation Impact
1.61

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article

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

Jinlei Liu, Yubing He, Han Cheng, Zhiqing Zhan et al.
2 citations
Rice
CRISPR and Genetic Engineering
1.61
article

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

Jinlei Liu, Yubing He, Han Cheng, Zhiqing Zhan, Dake Zhao, Yuwei Fu, Jinxin Liu, Hong Chen, Zhou Jiankun, Wen‐Qiang Chen, Ming‐Wei Wu, Lijun Tang, Chunming Liu, Tao Yang, Yiming Wang
article en
2 citations

Abstract

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.

RiceVol. 18(1)
Nanjing Agricultural University (CN), Yunnan University (CN), Beijing Botanical Garden (CN), Institute of Botany (CN), Institute of Crop Sciences (CN), University of Chinese Academy of Sciences (CN)
National Natural Science Foundation of China, Chinese Academy of Agricultural Sciences
Openalex Percentile: Top 15%
CRISPR and Genetic Engineering
1.61
DOIOpen Access PDFOpenAlex
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