Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes
Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes
复制标题
工程化生物可控 RNA 病毒载体，用于跨作物物种和基因型的非转基因基因组编辑
DOI：
10.1016/j.molp.2023.02.003
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发表时间：
2023-03-06
期刊：
影响因子：
27.5
通讯作者：
Li, Zhenghe
中科院分区：
文献类型：
--
作者：
Liu, Qian;Zhao, Chenglu;Li, Zhenghe
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.
