首页|Efficient and heritable A-to-K base editing in rice and tomato

Efficient and heritable A-to-K base editing in rice and tomato

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Cytosine and adenosine base editors(CBE and ABE)have been widely used in plants,greatly accelerating gene function research and crop breeding.Current base editors can achieve efficient A-to-G and C-to-T/G/A editing.However,efficient and heritable A-to-Y(A-to-T/C)editing remains to be developed in plants.In this study,a series of A-to-K base editor(AKBE)systems were constructed for monocot and dicot plants.Furthermore,nSpCas9 was replaced with the PAM-less Cas9 variant(nSpRY)to expand the target range of the AKBEs.Analysis of 228 T0 rice plants and 121 T0 tomato plants edited using AKBEs at 18 endogenous loci revealed that,in addition to highly efficient A-to-G substitution(41.0％on average),the plant AKBEs can achieve A-to-T conversion with efficiencies of up to 25.9 and 10.5％in rice and tomato,respectively.Moreover,the rice-optimized AKBE generates A-to-C conversion in rice,with an average efficiency of 1.8％,revealing the significant value of plant-optimized AKBE in creating genetic diversity.Although most of the A-to-T and A-to-C edits were chimeric,desired editing types could be transmitted to the T1 offspring,similar to the edits generated by the traditional ABE8e.Besides,using AKBEs to target tyrosine(Y,TAT)or cysteine(C,TGT)achieved the introduction of an early stop codon(TAG/TAA/TGA)of target genes,demonstrating its potential use in gene disruption.

Xinbo Li、Jiyong Xie、Chao Dong、Zai Zheng、Rundong Shen、Xuesong Cao、Xiaoyan Chen、Mugui Wang、Jian-Kang Zhu、Yifu Tian

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Ministry of Agriculture and Rural Affairs Key Laboratory of Gene Editing Technologies(Hainan),Institute of Crop Sciences and National Nanfan Research Institute,Chinese Academy of Agricultural Sciences,Sanya,Hainan 572024,China

Hainan Yazhou Bay Seed Lab,Sanya,Hainan 572024,China

Shanghai Center for Plant Stress Biology,CAS Center for Excellence in Molecular Plant Sciences,Chinese Academy of Sciences,Shanghai 201602,China

University of Chinese Academy of Sciences,Beijing 100049,China

Institute of Advanced Biotechnology,and School of Life Sciences,Southern University of Science and Technology,Shenzhen 518055,China

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中国博士后科学基金中国博士后科学基金Hainan Yazhou Bay Seed LaboratoryNanfan special project,CAAS国家自然科学基金

BX202200982022M720973B22C1000PZDXM231432188102

2024

10.1093/hr/uhad250

园艺研究(英文)

园艺研究(英文)

CSTPCD
ISSN：
年,卷(期)：2024.11(1)
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