Assessment of Miniature AsCas12f1 Variants for Gene Editing and Activation

IF 3.5 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology and Bioengineering Pub Date : 2025-03-19 DOI:10.1002/bit.28978

Chuanhong Ren, Zehua Bao

引用次数: 0

Abstract

Miniature CRISPR/Cas systems possess delivery advantages for gene therapy. The type V-F Cas12f1 from Acidibacillus sulfuroxidans is exceptionally compact (422 amino acids) and has been engineered by several studies as compact genome editing tools through protein and single guide RNA (sgRNA) engineering. However, a comparative evaluation of gene editing and activation efficiencies mediated by different AsCas12f1 variants and sgRNA scaffolds is lacking. This study tested combinations of four AsCas12f1 protein variants and six sgRNA scaffolds for their gene editing and transcription activation efficiencies. The protein variant AsCas12f1-HKRA performed the best in gene editing and activation when paired with sgRNA-en_v2.1 scaffold. Furthermore, we validated a super miniature gene activator by fusing a small activation domain to AsCas12f1-HKRA. Our findings recommend using AsCas12f1-HKRA and sgRNA-en_v2.1 for gene editing and activation applications.

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评估用于基因编辑和激活的微型 AsCas12f1 变体

微型CRISPR/Cas系统具有基因治疗的递送优势。来自硫酸氧化酸杆菌的V-F Cas12f1型非常紧凑（422个氨基酸），并通过蛋白质和单导RNA （sgRNA）工程被一些研究设计为紧凑的基因组编辑工具。然而，缺乏对不同AsCas12f1变异体和sgRNA支架介导的基因编辑和激活效率的比较评估。本研究测试了四种AsCas12f1蛋白变体和六种sgRNA支架的基因编辑和转录激活效率。当与sgRNA-en_v2.1支架配对时，蛋白变体AsCas12f1-HKRA在基因编辑和激活方面表现最好。此外，我们通过将一个小的激活域融合到AsCas12f1-HKRA上，验证了一个超微型基因激活子。我们的研究结果推荐使用AsCas12f1-HKRA和sgRNA-en_v2.1进行基因编辑和激活应用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology and Bioengineering 工程技术-生物工程与应用微生物

CiteScore

7.90

自引率

5.30%

发文量

280

审稿时长

2.1 months

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