清脆的
基因组编辑
计算生物学
计算机科学
生物
芯(光纤)
遗传学
基因
电信
作者
Hui Wang,Jian Ding,Jingyan Zhu,Xiaoshuang Liu,Ruihua Xu,Ruiying Qin,Danan Gu,Min Li,Pengcheng Wei,Juan Li
出处
期刊:aBIOTECH
[Springer Nature]
日期:2024-04-22
标识
DOI:10.1007/s42994-024-00157-5
摘要
Abstract Small mutations in the core promoter region of a gene may result in substantial changes in expression strengths. However, targeting TA-rich sequences of core promoters may pose a challenge for Cas9 variants such as SpCas9 and other G-rich PAM-compatible Cas9s. In this study, we engineered a unique FrCas9 system derived from Faecalibaculum rodentium for plant genome editing. Our findings indicate that this system is efficient in rice when the TATA sequence is used as a PAM. In addition, FrCas9 demonstrated activity against all 16 possible NNTA PAMs, achieving an efficiency of up to 35.3% in calli and generating homozygous or biallelic mutations in 31.3% of the T 0 transgenic plants. A proof-of-concept experiment to examine editing of the rice WX core promoter confirmed that FrCas9-induced mutations could modify gene expression and amylose content. Multiplex mutations and deletions were produced by bidirectional editing, mediated by FrCas9, using a single palindromic TATA sequence as a PAM. Moreover, we developed FrCas9-derived base editors capable of programmable conversion between A·T and G·C pairs in plants. This study highlights a versatile FrCas9 toolset for plant core promoter editing, offering great potential for the fine-tuning of gene expression and creating of new germplasms.
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