Megabase-scale human genome rearrangement with programmable bridge recombinases

IF 45.8 1区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Science Pub Date : 2025-09-25 DOI:10.1126/science.adz0276

Nicholas T. Perry, Liam J. Bartie, Dhruva Katrekar, Gabriel A. Gonzalez, Matthew G. Durrant, James J. Pai, Alison Fanton, Juliana Q. Martins, Masahiro Hiraizumi, Chiara Ricci-Tam, Hiroshi Nishimasu, Silvana Konermann, Patrick D. Hsu

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引用次数: 0

Abstract

Bridge recombinases are naturally occurring RNA-guided DNA recombinases that we previously demonstrated can programmably insert, excise, and invert DNA in vitro and in Escherichia coli . In this study, we report the discovery and engineering of the bridge recombinase ortholog ISCro4 for universal rearrangements of the human genome. We defined strategies for the optimal application of bridge systems, leveraging mechanistic insights to improve their targeting specificity. Through rational engineering of the ISCro4 bridge RNA and deep mutational scanning of its recombinase, we achieved up to 20% insertion efficiency into the human genome and genome-wide specificity as high as 82%. We further demonstrated intrachromosomal inversion and excision, mobilizing up to 0.93 megabases of DNA. Lastly, we provided proof-of-concept for plasmid-based excision of disease-relevant gene regulatory regions or repeat expansions.

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用可编程桥式重组酶进行百万级人类基因组重排

桥接重组酶是天然存在的rna引导的DNA重组酶，我们之前已经证明它可以在体外和大肠杆菌中可编程地插入、切除和转化DNA。在这项研究中，我们报道了桥梁重组酶同源物ISCro4的发现和工程，用于人类基因组的普遍重排。我们定义了桥梁系统的最佳应用策略，利用机制的见解来提高其靶向特异性。通过对ISCro4桥接RNA进行合理的工程设计，并对其重组酶进行深度突变扫描，我们实现了高达20%的插入效率和高达82%的全基因组特异性。我们进一步展示了染色体内反转和切除，动员高达0.93兆碱基的DNA。最后，我们提供了基于质粒的疾病相关基因调控区域切除或重复扩增的概念证明。

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

Science 综合性期刊-综合性期刊

CiteScore

61.10

自引率

0.90%

发文量

审稿时长

2.1 months

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