Fusing T5 exonuclease with Cas9 and Cas12a increases the frequency and size of deletion at target sites

查看全文 作　　者：Qianwei [1,2]Zhang;Kangquan [1]Yin;Guanwen [1,2]Liu;Shengnan [1]Li;Mengou [1,2]Li;Jin-Long [1,2]Qiu

机构地区：[1]State Key Laboratory of Plant Genomics,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China;[2]University of Chinese Academy of Sciences,Beijing 100049,China

出　　处：《Science China(Life Sciences)》索引2020年第63卷第12期,共10页

基　　金：supported by grants from the National Transgenic Science and Technology Program of China(2019ZX08010-003,2019ZX08010-001,2018ZX0801002B);the National Key Research and Development Program of China(2016YFD0100602);the Scientific Program of Beijing Municipal Commission of Science and Technology(Z171100001517001);the National Natural Science Foundation of China(31672015)to J.L.Q。 摘　　要：CRISPR/Cas systems, especially CRISPR/Cas9, generally result in small insertions/deletions, which are unlikely to eliminate the functions of regulatory and other non-coding sequences. To generate larger genomic deletions usually requires the use of pairs of guide RNAs. Here we show that it is possible to create such deletions with a single guide RNA by fusing Cas9 or Cas12a with T5 exonuclease(T5exo). These fusion constructs were found to increase both the frequency and size of deletions at target loci in rice protoplasts and seedlings. Moreover, the genome editing efficiencies of Cas9 and Cas12a were also enhanced by fusion with T5 exonuclease. These T5exo-Cas fusions expand the CRISPR toolbox, and facilitate knockout of regulatory and non-coding DNA sequences. From a wider standpoint, our results suggest a general strategy for producing larger deletions using other Cas nucleases. 关键词：Cas9 Cas12a T5 exonuclease genome editing DELETION

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Fusing T5 exonuclease with Cas9 and Cas12a increases the frequency and size of deletion at target sites

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