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通过直接选在哺乳动物细胞中获得的可切换光的转录因子

Liyuan Zhu¹, Harold M McNamara^2,3, Jared E Toettcher⁴

¹Department of Chemistry, Princeton University, Princeton, NJ, 08544, USA.

Nature communications

|June 2, 2023

概括

研究人员开发了一种新方法,用于光遗传学创建可切换光的蛋白质. 这种技术可以发现具有光控制功能的新型蛋白质变体,从而推动生物研究.

科学领域:

生物化学生物化学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

光遗传工具通过使用光来精确控制生物过程.
开发具有可预测功能的新的可切换光的蛋白质变体是一个重大挑战.
目前缺乏工程或发现这种变体的一般策略.

研究的目的:

开发一种可通用的方法,用于直接在哺乳动物细胞中设计和发现新的光遗传工具.
为了创建可切换光的蛋白质变体,以加强对生物功能的控制.
为了简化单蛋白光遗传开关的识别.

主要方法:

适应蛋白质域插入和哺乳动物细胞表达策略.
通过插入AsLOV2可光切换域来生成和选候选光遗传工具库.
在哺乳动物细胞中表现出可光切换活性的变体中选择明暗.

主要成果:

使用Gal4-VP64转录因子演示了该方法,创建了LightsOut转录因子.
在黑暗和蓝光条件之间实现了转录活性>150倍的变化.
向其他指域 (Cys6Zn2和C2H2) 展示了可切换光功能的概括.

结论:

开发的方法有效地在哺乳动物细胞中产生和选光遗传工具.
这种方法有助于发现单蛋白光遗传开关,即使先前知识有限.
这些发现为光遗传学调节更广泛的转录因子提供了基础.

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