利用增强型 CRISPR-Cas12f 系统高效特异地调控基因表达。

IF 4.6 3区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Yeounsun Oh, Lee Wha Gwon, Hyomin K. Lee, Junho K. Hur, Kwang-Hyun Park, Kee-Pyo Kim, Seung Hwan Lee
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引用次数: 0

摘要

最近开发的 CRISPR 激活器(CRISPRa)系统利用基于 CRISPR-Cas 效应器的转录激活器,在不造成 DNA 损伤的情况下有效控制目标基因的表达。然而,由于 CRISPR-Cas 模块本身的局限性,现有的基于 Cas9/Cas12a 的 CRISPRa 系统需要在功效和准确性方面进行改进。为了克服这些局限性,有效、准确地调控基因表达,我们开发了一种基于小型CRISPR-Cas效应子Candidatus Woesearchaeota Cas12f(CWCas12f)的高效CRISPRa系统。与现有的CRISPRa系统相比,通过对CRISPR-Cas模块进行工程化、连接激活结构域以及使用不同的连接体和核定位信号序列组合,优化后的eCWCas12f-VPR系统能够有效地对基因表达进行靶标特异性调控。本研究开发的 eCWCas12f-VPR 系统在控制生物体内源基因转录方面具有巨大潜力,可作为未来基因治疗和生物研究的基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Highly efficient and specific regulation of gene expression using enhanced CRISPR-Cas12f system

Highly efficient and specific regulation of gene expression using enhanced CRISPR-Cas12f system

Highly efficient and specific regulation of gene expression using enhanced CRISPR-Cas12f system
The recently developed CRISPR activator (CRISPRa) system uses a CRISPR-Cas effector-based transcriptional activator to effectively control the expression of target genes without causing DNA damage. However, existing CRISPRa systems based on Cas9/Cas12a necessitate improvement in terms of efficacy and accuracy due to limitations associated with the CRISPR-Cas module itself. To overcome these limitations and effectively and accurately regulate gene expression, we developed an efficient CRISPRa system based on the small CRISPR-Cas effector Candidatus Woesearchaeota Cas12f (CWCas12f). By engineering the CRISPR-Cas module, linking activation domains, and using various combinations of linkers and nuclear localization signal sequences, the optimized eCWCas12f-VPR system enabled effective and target-specific regulation of gene expression compared with that using the existing CRISPRa system. The eCWCas12f-VPR system developed in this study has substantial potential for controlling the transcription of endogenous genes in living organisms and serves as a foundation for future gene therapy and biological research.
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来源期刊
Gene Therapy
Gene Therapy 医学-生化与分子生物学
CiteScore
9.70
自引率
2.00%
发文量
67
审稿时长
4-8 weeks
期刊介绍: Gene Therapy covers both the research and clinical applications of novel therapeutic techniques based on a genetic component. Over the last few decades, significant advances in technologies ranging from identifying novel genetic targets that cause disease through to clinical studies, which show therapeutic benefit, have elevated this multidisciplinary field to the forefront of modern medicine.
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