Engineering Miniature CRISPR-Cas Un1Cas12f1 for Efficient Base Editing

doi:10.1016/j.omtn.2024.102201

	Engineering Miniature CRISPR-Cas Un1Cas12f1 for Efficient Base Editing
	Hu, Yueer1 ; Han, Linxiao1 ; Mo, Qiqin1 ; Du, Zengming 2; Jiang, Wei 2; Wu, Xia 3; Zheng, Jing 2; Xiao, Xiao 2; Sun, Yadong4 ; Ma, Hanhui1
	2024
发表期刊	MOLECULAR THERAPY: NUCLEIC ACIDS (IF:6.5[JCR-2023],6.7[5-Year])
ISSN	2162-2531
卷号	35 期号:2
发表状态	已发表
DOI	10.1016/j.omtn.2024.102201
摘要	Adeno-associated virus (AAV) is a relatively safe and ef fi cient vector for gene therapy. However, due to its 4.7 -kb limit of cargo, SpCas9-mediated base editors cannot be packaged into a single AAV vector, which hinders their clinical application. The development of ef fi cient miniature base editors becomes an urgent need. Un1Cas12f1 is a class II V -F -type CRISPRCas protein with only 529 amino acids. Although Un1Cas12f1 has been engineered to be a base editor in mammalian cells, the base-editing ef fi ciency is less than 10%, which limits its therapeutic applications. Here, we developed hypercompact and high-ef fi ciency base editors by engineering Un1Cas12f1, fusing non-speci fi c DNA binding protein Sso7d, and truncating single guide RNA (sgRNA), termed STUminiBEs. We demonstrated robust A -to -G conversion (54% on average) by STUminiABEs or C -to -T conversion (45% on average) by STUminiCBEs. We packaged STUminiCBEs into AAVs and successfully introduced a premature stop codon on the PCSK9 gene in mammalian cells. In sum, STUminiBEs are ef fi cient miniature base editors and could readily be packaged into AAVs for biological research or biomedical applications.
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收录类别	SCI
语种	英语
资助项目	Shanghai Institute for Advanced Immunochemical Studies (SIAIS) at ShanghaiTech University - National Natural Science Foundation of China[31970591] ; Shanghai Science and Technology Innovation Action Plan[21JC1404800]
WOS研究方向	Research & Experimental Medicine
WOS类目	Medicine, Research & Experimental
WOS记录号	WOS:001239928800001
出版者	CELL PRESS
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/370088
专题	生命科学与技术学院_博士生生命科学与技术学院_PI研究组_马涵慧组生命科学与技术学院_PI研究组_孙亚东组
共同第一作者	Han, Linxiao
通讯作者	Ma, Hanhui
作者单位	1.Gene Editing Center, School of Life Science and Technology, ShanghaiTech University, Shanghai, China. 2.Belief BioMed (Shanghai) Inc. Shanghai, China. 3.School of Biotechnology, East China University of Science and Technology, Shanghai, China. 4.School of Life Science and Technology, ShanghaiTech University, Shanghai, China.
第一作者单位	生命科学与技术学院
通讯作者单位	生命科学与技术学院
第一作者的第一单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Hu, Yueer,Han, Linxiao,Mo, Qiqin,et al. Engineering Miniature CRISPR-Cas Un1Cas12f1 for Efficient Base Editing[J]. MOLECULAR THERAPY: NUCLEIC ACIDS,2024,35(2).
APA	Hu, Yueer.,Han, Linxiao.,Mo, Qiqin.,Du, Zengming.,Jiang, Wei.,...&Ma, Hanhui.(2024).Engineering Miniature CRISPR-Cas Un1Cas12f1 for Efficient Base Editing.MOLECULAR THERAPY: NUCLEIC ACIDS,35(2).
MLA	Hu, Yueer,et al."Engineering Miniature CRISPR-Cas Un1Cas12f1 for Efficient Base Editing".MOLECULAR THERAPY: NUCLEIC ACIDS 35.2(2024).