Robust CRISPR/Mb2Cas12a genome editing tools in cotton plants

IF 23.7 Q1 MICROBIOLOGY iMeta Pub Date : 2024-06-04 DOI:10.1002/imt2.209
Fengjiao Hui, Xu Tang, Bo Li, Muna Alariqi, Zhongping Xu, Qingying Meng, Yongxue Hu, Guanying Wang, Yong Zhang, Xianlong Zhang, Shuangxia Jin
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Abstract

The efficiency and accuracy of the CRISPR/Mb2Cas12a system were demonstrated in cotton, achieving an efficiency of over 90% at target sites. Notably, Mb2Cas12a exhibited significant tolerance under different temperatures ranging from 22°C to 32°C. Additionally, the Mb2Cas12a system revealed effective editing at more relaxed VTTV PAM sites in the cotton genome, which expanded the genome editing range by approximately 2.6-fold than the wide-type LbCas12a. Finally, a multiplex genome editing system was also developed based on Mb2Cas12a, enabling simultaneous editing of eight target sites using a single crRNA cassette.

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棉花植物中强大的 CRISPR/Mb2Cas12a 基因组编辑工具
CRISPR/Mb2Cas12a系统的效率和准确性在棉花中得到了验证,在目标位点的效率超过90%。值得注意的是,Mb2Cas12a在22°C至32°C的不同温度下表现出明显的耐受性。此外,Mb2Cas12a 系统还能有效编辑棉花基因组中更宽松的 VTTV PAM 位点,与宽型 LbCas12a 相比,基因组编辑范围扩大了约 2.6 倍。最后,基于 Mb2Cas12a 还开发了一种多重基因组编辑系统,可使用单个 crRNA 盒同时编辑八个目标位点。
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