Engineered CRISPR RNA improves the RNA cleavage efficiency of hfCas13X

IF 3.5 4区 生物学 Q1 Biochemistry, Genetics and Molecular Biology
Zehui Liu, Wenxia Zhang, Haili Wang, Pingping Shangguan, Tong Pan, Yimu Yang, Yi Zhang, Xi Mao, Yingle Liu, Qi Zhang
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Abstract

As the most compact variant in the Cas13 family, CRISPR-Cas13X holds considerable promise for gene therapy applications. The development of high-fidelity Cas13X (hfCas13X) mutants has enhanced the safety profile for in vivo applications. However, a notable reduction in on-target cleavage efficiency accompanies the diminished collateral cleavage activity in hfCas13X. In this study, we obtained two engineered crRNA mutants that notably enhance the on-target cleavage efficiency of hfCas13X. Furthermore, we have identified a novel crRNA structure that consistently augments the on-target cleavage efficiency of hfCas13X across various cellular environments, without significant enhancement of its collateral activity. These findings collectively enrich the gene-editing toolkit, presenting a more effective hfCas13X system for future research and application.

Abstract Image

设计的 CRISPR RNA 提高了 hfCas13X 的 RNA 切割效率。
作为Cas13家族中最紧凑的变体,CRISPR-Cas13X在基因治疗应用中大有可为。高保真 Cas13X(hfCas13X)突变体的开发提高了体内应用的安全性。然而,伴随着 hfCas13X 附带裂解活性的降低,靶上裂解效率也明显降低。在这项研究中,我们获得了两种工程化的 crRNA 突变体,它们能显著提高 hfCas13X 的靶向切割效率。此外,我们还发现了一种新型的 crRNA 结构,它能在各种细胞环境中持续提高 hfCas13X 的靶上切割效率,而不会显著增强其附带活性。这些发现共同丰富了基因编辑工具包,为未来的研究和应用提供了更有效的 hfCas13X 系统。
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
FEBS Letters
FEBS Letters 生物-生化与分子生物学
CiteScore
7.00
自引率
2.90%
发文量
303
审稿时长
1.0 months
期刊介绍: FEBS Letters is one of the world''s leading journals in molecular biology and is renowned both for its quality of content and speed of production. Bringing together the most important developments in the molecular biosciences, FEBS Letters provides an international forum for Minireviews, Research Letters and Hypotheses that merit urgent publication.
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