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Improving gene editing of CRISPR/Cas9 using the callus-specific promoter pYCE1 in cassava (Manihot esculenta Crantz)

文献类型: 外文期刊

作者: Li, Yuanchao 1 ; Bao, Ruxue 2 ; Li, Mengtao 2 ; Zeng, Changying 3 ; Yang, Haojie 2 ; Yao, Yuan 2 ; Li, Youzhi 1 ; Wang, Wenquan 3 ; Chen, Xin 2 ;

作者机构: 1.Guangxi Univ, Coll Life Sci & Technol, State Key Lab Conservat & Utilizat Subtrop Agrobio, Nanning, Guangxi, Peoples R China

2.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Natl Key Lab Trop Crop Breeding, Sanya, Hainan, Peoples R China

3.Hainan Univ, Inst Trop Agr & Forestry, Haikou, Peoples R China

4.Hainan Inst Trop Agr Resources, Key Lab Biol & Genet Resources Trop Crops Hainan P, Haikou, Hainan, Peoples R China

5.Chinese Acad Trop Agr Sci, Sanya Res Inst, Sanya, Hainan, Peoples R China

关键词: cassava; callus; promoter; gene editing; target

期刊名称:FRONTIERS IN PLANT SCIENCE ( 影响因子:4.8; 五年影响因子:5.7 )

ISSN: 1664-462X

年卷期: 2025 年 16 卷

页码:

收录情况: SCI

摘要: Previous studies have demonstrated that an appropriate promoter can drive Cas9 transcription in the CRISPR/Cas9 system, which improves the efficiency of gene editing. Here, we identified and characterized callus-specific promoters to enhance gene editing efficiency in cassava. From the transcriptome data of 11 cassava tissues, the gene named YCE1 was identified to exhibit callus-specific expression. Its promoter (pYCE1) could efficiently and specifically drive EGFP transcription in callus tissues. Given that friable embryogenic callus (FECs) is the recipient for genetic transformation in cassava, we replaced the original 35S promoter with pYCE1 to drive Cas9 transcription for improving the CRISPR/Cas9 gene editing system. In single-gene editing, the mutation rate was significantly increased, which reached an overall mutation rate of 95.24% and a homozygous mutation rate of 52.38%, compared with 62.07% and 37.93% with the 35S promoter, respectively. Furthermore, achieving a dual-gene homozygous mutation rate of 64.71% in dual-gene editing demonstrated the high efficiency of pYCE1 in the gene editing application for cassava. These results underscore the potential of pYCE1 to enhance gene editing efficiency in the CRISPR/Cas9 system of cassava. This approach paves the way for advanced gene function research and genetic breeding in cassava.

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