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Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes

摘要CRISPR/Cas genome-editing tools provide unprecedented opportunities for basic plant biology research and crop breeding.However,the lack of robust delivery methods has limited the widespread adoption of these revolutionary technologies in plant science.Here,we report an efficient,non-transgenic CRISPR/Cas delivery platform based on the engineered tomato spotted wilt virus(TSWV),an RNA virus with a host range of over 1000 plant species.We eliminated viral elements essential for insect transmission to liberate genome space for accommodating large genetic cargoes without sacrificing the ability to infect plant hosts.The resulting non-insect-transmissible viral vectors enabled effective and stable in planta de-livery of Cas12a and Cas9 nucleases as well as adenine and cytosine base editors.In systemically infected plant tissues,the deconstructed TSWV-derived vectors induced efficient somatic gene mutations and base conversions in multiple crop species with little genotype dependency.Plants with heritable,bi-allelic mutations could be readily regenerated by culturing the virus-infected tissues in vitro without antibiotic se-lection.Moreover,we showed that antiviral treatment with ribavirin during tissue culture cleared the viral vectors in 100%of regenerated plants and further augmented the recovery of heritable mutations.Because many plants are recalcitrant to stable transformation,the viral delivery system developed in this work pro-vides a promising tool to overcome gene delivery bottlenecks for genome editing in various crop species and elite varieties.

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作者 Qian Liu [1] Chenglu Zhao [1] Kai Sun [1] Yinlu Deng [1] Zhenghe Li [2] 学术成果认领
作者单位 State Key Laboratory of Rice Biology,Institute of Biotechnology,Zhejiang University,Hangzhou,China [1] State Key Laboratory of Rice Biology,Institute of Biotechnology,Zhejiang University,Hangzhou,China;Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insect Pests,Zhejiang University,Hangzhou,China;Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province,Zhejiang University,Hangzhou,China [2]
DOI 10.1016/j.molp.2023.02.003
发布时间 2024-04-09(万方平台首次上网日期,不代表论文的发表时间)
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分子植物(英文版)

分子植物(英文版)

2023年3期

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