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Boosting genome editing in plants with single transcript unit surrogate reporter systems

Plant Communications 2024年第6期5卷 32-43页

作者： Xu Tang Qiurong Ren Xiaodan Yan Rui Zhang Li Liu Qinqin Han Xuelian Zheng Yiping Qi Hongyuan Song Yong Zhang Chongqing Key Laboratory of Plant Resource Conservation and Germplasm Innovation Integrative Science Center of Germplasm Creation in Western China(Chongqing)Science CitySchool of Life SciencesSouthwest UniversityChongqing 400715China Key Laboratory of Agricultural Biosafety and Green Production of Upper Yangtze River Ministry of EducationChongqing 400715China College of Horticulture and Landscape Architecture Southwest UniversityChongqing 400715China Department of Biotechnology School of Life Sciences and TechnologyCenter for Informational BiologyUniversity of Electronic Science and Technology of ChinaChengdu 610054China School of Synbiology School of Life ScienceShanxi UniversityTaiyuan 030006China Department of Plant Science and Landscape Architecture University of MarylandCollege ParkMD 20742USA Institute for Bioscience and Biotechnology Research University of MarylandRockvilleMD 20850USA

CRISPR-Cas-based genome editing holds immense promise for advancing plant genomics and crop ***,the challenge of low editing activity complicates the identification of editing *** this study,we introduce multiple single transcript unit surrogate reporter(STU-SR)systems to enhance the selection of genome-edited *** systems use the same single guide RNAs designed for endogenous genes to edit reporter genes,establishing a direct link between reporter gene editing activity and that of endogenous *** strategies are used to restore functional reporter genes after genome editing,including efficient single-strand annealing(SSA)for homologous recombination in STUSR-SSA ***-SR-base editor systems leverage base editing to reinstate the start codon,enriching C-to-T and A-to-G base editing *** results showcase the effectiveness of these STU-SR systems in enhancing genome editing events in the monocot rice,encompassing Cas9 nuclease-based targeted mutagenesis,cytosine base editing,and adenine base *** systems exhibit compatibility with Cas9 variants,such as the PAM-less SpRY,and are shown to boost genome editing in Brassica oleracea,a dicot vegetable *** summary,we have developed highly efficient and versatile STU-SR systems for enrichment of genome-edited plants.

关键词： CRISPR-Cas9 single transcript unit STU surrogate reporter SR single strand annealing SSA cyto-sine base editing adenine base editing

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