加州大学Jennifer A. Doudna团队取得一项新突破。他们提出了最小RNA-引导核酸酶的结构和进化引导设计。2026年7月16日出版的《科学》发表了这项成果。

研究组提出了一种蛋白质设计策略，将结构引导的反折叠模型与进化信息残基约束结合起来，生成TnpB的活性、分化变体，TnpB是一种最小的CRISPR-Cas12样核酸酶，称为SynTnpBs。对人工智能生成的变体进行高通量筛选，产生的编辑器在细菌、植物和人类细胞中保留或超过了野生型活性。基于低温电子显微镜的结构测定显示了RNA-DNA界面在不同构象之间的稳定接触，证明了这种方法的设计潜力。总之，这些结果建立了创建非天然RNA引导核酸酶和构象活性核酸结合物的策略，扩大了可设计的蛋白质空间。

研究人员表示，设计具有不受进化限制的特性的RNA引导核酸酶可以扩展可编程基因组编辑能力。然而，产生具有机械酶特性的多种多结构域蛋白仍然具有挑战性。

附：英文原文

Title: Structure and evolution-guided design of minimal RNA-guided nucleases

Author: Petr Skopintsev, Isabel Esain-Garcia, Evan C. DeTurk, Peter H. Yoon, Zehan Zhou, Trevor Weiss, Maris Kamalu, Ajit Chamraj, Kenneth J. Loi, Conner J. Langeberg, Ron S. Boger, Hunter Nisonoff, Hannah M. Karp, Lin-Xing Chen, Honglue Shi, Kamakshi Vohra, Jillian F. Banfield, Jamie H. D. Cate, Steven E. Jacobsen, Jennifer A. Doudna

Issue&Volume: 2026-07-16

Abstract: The design of RNA-guided nucleases with properties not limited by evolution can expand programmable genome-editing capabilities. However, generating diverse multidomain proteins with robust enzymatic properties remains challenging. Here, we use a protein design strategy that couples a structure-guided inverse-folding model with evolution-informed residue constraints to generate active, divergent variants of TnpB, a minimal CRISPR-Cas12–like nuclease, termed SynTnpBs. High-throughput screening of artificial intelligence–generated variants yielded editors that retained or exceeded wild-type activity in bacterial, plant, and human cells. Cryo–electron microscopy–based structure determination of the most divergent variant revealed stabilizing contacts in the RNA–DNA interfaces across conformations, demonstrating the design potential of this approach. Together, these results establish a strategy for creating non-natural RNA-guided nucleases and conformationally active nucleic acid binders, enlarging the designable protein space.

DOI: aed6123

Source: https://www.science.org/doi/10.1126/science.aed6123

期刊信息

Science：《科学》，创刊于1880年。隶属于美国科学促进会，最新IF：63.714

官方网址：https://www.sciencemag.org/

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