设计抗CRISPR蛋白质,以创建CRISPR-Cas蛋白开关,用于可激活的基因组编辑和病毒蛋白酶检测
Wenyuan Kang1,2, Fei Xiao1, Xi Zhu3
1State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan Provincial Key Laboratory of Biomacromolecular Chemical Biology, Hunan University, Changsha, 410082, P. R. China.
Angewandte Chemie (International ed. in English)
|February 26, 2024
概括
研究人员使用抗CRISPR蛋白质设计了合成的CRISPR-Cas开关. 这些开关可以精确控制Cas12a的活动,促进蛋白酶诱导基因组编辑和敏感的病毒诊断.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 合成生物学 合成生物学
背景情况:
- 在活性状态之间切换的蛋白质对于生物感知和控制至关重要.
- 可激活的CRISPR-Cas系统提供精确的基因操纵和诊断,但缺乏直接的功能控制.
- 反CRISPR蛋白质被探索为创建可控制的Cas系统的组件.
研究的目的:
- 通过使用抗CRISPR蛋白来进行受控的Cas12a激活,设计合成Cas蛋白开关 (CasPSs).
- 开发能够响应特定的蛋白质溶解事件的CasPS,用于针对性基因编辑和诊断.
- 为合理的蛋白质接口工程利用计算化学.
主要方法:
- 计算化学技术包括分子指纹分析,静电电位映射和结合自由能计算.
- 在Cas12a和抗CRISPR蛋白质 (AcrVA4,AcrVA5) 之间的接口的合理工程.
- 对直角蛋白质酶响应的CasPSs的生成和表征.
主要成果:
- 成功设计了将蛋白质溶解事件转化为高切换率 (高达34.3倍) 的Cas12a激活的CasPSs.
- 在使用开发的CasPSs的哺乳动物细胞中,证明了可诱导蛋白解的基因组编辑.
- 在感染期间展示了病毒蛋白酶活动的敏感检测.
结论:
- 开发了一种新的策略,用于创建基于抗CRISPR蛋白质工程的合成CRISPR-Cas开关.
- CasPSs可以精确控制CRISPR-Cas功能,从而在基因操纵和临床诊断中实现新的应用.
- 这种方法为推进可控制的基因编辑工具和诊断方法提供了一个有希望的平台.
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