数字农科院2.0

Engineering a robust Cas12i3 variant-mediated wheat genome editing system

文献类型： 外文期刊

作者： Wang, Wenxue;Yan, Lei;Li, Jingying;Zhang, Chen;He, Yubing;Li, Shaoya;Xia, Lanqin

作者机构：

关键词： wheat (Triticum aestivum L.);CRISPR/Cas12i3;genome editing;T5 Exonuclease (T5E)

期刊名称： PLANT BIOTECHNOLOGY JOURNAL

ISSN： 1467-7644

年卷期： 2024 年

页码：

收录情况： SCIE（2024版）

摘要： Wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD) is one of the most important food crops in the world. CRISPR/Cas12i3, which belongs to the type V-I Cas system, has attracted extensive attention recently due to its smaller protein size and its less-restricted canonical 'TTN' protospacer adjacent motif (PAM). However, due to its relatively lower editing efficacy in plants and the hexaploidy complex nature of wheat, Cas12i3/Cas12i3-5M-mediated genome editing in wheat has not been documented yet. Here, we report the engineering of a robust Cas12i3-5M-mediated genome editing system in wheat through the fusion of T5 exonuclease (T5E) in combination with an optimised crRNA expression strategy (Opt). We first showed that fusion of T5E, rather than ExoI, to Cas12i3-5M increased the gene editing efficiencies by up to 1.34-fold and 3.87-fold, compared to Cas12i3-5M and Cas12i3 in HEK293T cells, respectively. However, its editing efficiency remains low in wheat. We then optimised the crRNA expression strategy and demonstrated that Opt-T5E-Cas12i3-5M could enhance the editing efficiency by 1.20- to 1.33-fold and 4.05- to 7.95-fold in wheat stable lines compared to Opt-Cas12i3-5M and Opt-Cas12i3, respectively, due to progressive 5 '-end resection of the DNA strand at the cleavage site with increased deletion size. The Opt-T5E-Cas12i3-5M enabled an editing efficiency ranging from 60.71% to 90.00% across four endogenous target genes in stable lines of three elite Chinese wheat varieties. Together, the developed robust Opt-T5E-Cas12i3-5M system enriches wheat genome editing toolkits for either biological research or genetic improvement and may be extended to other important polyploidy crop species.

分类号：

  • 相关文献

[1]Present and future prospects for wheat improvement through genome editing and advanced technologies. Shaoya Li,Chen Zhang,Jingying Li,Lei Yan,Ning Wang,Lanqin Xia. 2021

[2]Genome editing in rice using CRISPR/Cas12i3. Lv, Ping,Su, Fei,Chen, Fangyuan,Yan, Chunxue,Xia, Dandan,Sun, Hui,Li, Shanshan,Duan, Zhiqiang,Ma, Changle,Zhang, Hui,Wang, Mugui,Niu, Xiaomu,Zhu, Jian-Kang,Zhang, Jinshan. 2023

[3]The Major Factors Causing The Microspore Abortion Of Genic Male Sterile Mutant Nwms1 In Wheat (Triticum Aestivum L.). Xu, Qiaoqiao,Jiao, Zhixin,An, Junhang,Li, Junchang,Zhang, Jing,Li, Huijuan,Li, Qiaoyun,Niu, Hao,Niu, Jishan,Jiang, Yumei. 2019

[4]HapIII of TaSAP1-A1, a Positively Selected Haplotype in Wheat Breeding. Chang Jian-zhong,Hao Chen-yang,Chang Xiao-ping,Zhang Xue-yong,Jing Rui-lian. 2014

[5]Factorial cross analysis of pre-harvest sprouting resistance in white wheat. Jiang, GL,Xiao, SH. 2005

[6]Mapping QTLs for stomatal density and size under drought stress in wheat (Triticum aestivum L.). Wang Shu-guang,Jia Shou-shan,Sun Dai-zhen,Fan Hua,Chang Xiao-ping,Jing Rui-lian. 2016

[7]Development of near-infrared reflectance spectroscopy models for quantitative determination of water-soluble carbohydrate content in wheat stem and glume. Wang, Zhenghang,Liu, Xiulin,Chang, Xiaoping,Jing, Ruilian,Wang, Zhenghang,Liu, Xiulin,Li, Runzhi.

[8]Length of internode and spike: how do they contribute to plant height of wheat at an individual QTL level?. Wang, L.,Cui, F.,Ding, A. M.,Li, J.,Zhao, C. H.,Li, X. F.,Feng, D. S.,Wang, H. G.,Wang, L.,Wang, J. P.,Cui, F.,Ding, A. M.,Li, J..

[9]Double-stranded RNA in the biological control of grain aphid (Sitobion avenae F.). Wang, Dahai,Sun, Yongwei,Wang, Hui,Xia, Lanqin,Wang, Dahai,Liu, Qi,Li, Xia.

[10]The Mirna-Mrna Networks Involving Abnormal E.nergy And Hormone Metabolisms R estrict Tillering In A Wheat Mutant Dmc. Ni, Yongjing,He, Ruishi,Jiao, Zhixin,An, Junhang,Zheng, Yongxing,Jiang, Yumei,Li, Huijuan,Li, Junchang,Niu, Jishan,Niu, Hao,Li, Qiaoyun,Zhang, Jing. 2019

[11]Allelic Variation in Loci for Adaptive Response and Its Effect on Agronomical Traits in Chinese Wheat (Triticum aestivum L.). Gao Li-feng,Liu Pan,Gu Yan-chun,Jia Ji-zeng. 2014

[12]Wheat traits and the associated loci conferring radiation use efficiency. Li Y., Tao F., Hao Y., Tong J., Xiao Y., He Z., Reynolds M.. 2022

[13]Resistance Performance Of Wheat Stripe Rust Resistance Gene Yr41 And Its Effect On Yield Parameters In F-2 Populations Under Field Conditions. Jiang, C, Wang, XS, Chen, WQ, Liu, TG, Zhong, SF, Huang, QL, Ren, TH, Li, Z, Tan, FQ, Luo, PG. 2020

[14]Quantitative trait loci mapping and candidate gene analysis of stoma-related traits in wheat (Triticum aestivum L.) glumes. Ning Li,Fanfan Dong,Tongtong Liu,Jinwen Yang,Yugang Shi,Shuguang Wang,Daizhen Sun,Ruilian Jing. 2022

[15]Genome-wide identification of Glutathione peroxidase (GPX) family genes and silencing TaGPX3.2A reduced disease resistance in wheat. Baihui Jiang,Chang Su,Youning Wang,Xiao Xu,Yan Li,Dongfang Ma. 2023

[16]Assessing genetic diversity for some Pakistani bread wheat (Triticum aestivum L.) genotypes under drought stress based on yield traits. Khan, Mueen Alam,Yousaf, Malik Waqar,Ahmed, Hafiz Ghulam Muhu-Din,Fatima, Noor,Alam, Beena. 2024

[17]Mapping QTLs for adult-plant resistance to powdery mildew and stripe rust using a recombinant inbred line population derived from cross Qingxinmai × 041133. Yahui Li,Jinghuang Hu,Huailong Lin,Dan Qiu,Yunfeng Qu,Jiuyuan Du,Lu Hou,Lin Ma,Qiuhong Wu,Zhiyong Liu,Yijun Zhou,Hongjie Li. 2024

[18]TaWRKY24 integrates the tryptophan metabolism pathways to participate in defense against Fusarium crown rot in wheat. Xu, Xing,Yu, Tai-Fei,Wei, Ji-Tong,Ma, Xiao-Fei,Liu, Yong-Wei,Zhang, Jin-Peng,Zheng, Lei,Hou, Ze-Hao,Chen, Jun,Zhou, Yong-Bin,Chen, Ming,Ma, Jian,Jiang, Yun-Feng,Ji, Hu-Tai,Li, Li-Hui,Ma, You-Zhi,Zhang, Zhi-An,Xu, Zhao-Shi. 2024

[19]TaPUB57 confers drought tolerance, governs grain size and salt sensitivity by ubiquitinating TaEXPB3 in rice. Chunmei Yu,Min Wang,Long Li,Yuying Li,Yanfei Zhang,Jie Zhang,Xin Chen,Lili Yang,Chaonan Li,Jingyi Wang,Guangchao Liu,Matthew P. Reynolds,Qi Xie,Ruilian Jing,Xinguo Mao. 2025

[20]Crispr-S: An Active Interference Element F.or A Rapid And I nexpensive Selection Of Genome-Edited, Transgene-Free Rice Plants. Huang, Jian-Zhong,Huang, Jian-Zhong,Liu, Song-Mei,Shu, Qing-Yao,Tan, Yuan-Yuan,Lu, Hai-Ping,Lu, Hai-Ping,Chen, Wu-Yang,Shu, Qing-Yao,Tan, Yuan-Yuan,Chen, Wu-Yang,Shu, Qing-Yao,Huang, Jian-Zhong,Xu, Shou-Ling,Zhou, Xin. 2017

作者其他论文 更多>>