大规模评估RNA结合蛋白激活外显子纳入的能力
Jonathan C Schmok1,2,3,4, Manya Jain1,2,3, Lena A Street5
1Department of Cellular and Molecular Medicine, University of California San Diego, La Jolla, CA, USA.
Nature biotechnology
|January 3, 2024
概括
研究人员发现了新的RNA结合蛋白 (RBPs),通过新型的记者系统控制基因剪接. 这一发现为精确控制拼接提供了新的工具,影响基因表达和细胞功能.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物化学 生化学
背景情况:
- RNA结合蛋白 (RBPs) 通过控制替代拼接来调节基因表达.
- 了解哪些RBPs直接影响外子包容对于破译细胞生存机制至关重要.
研究的目的:
- 识别直接促进替代外显子纳入的新型RNA结合蛋白 (RBPs).
- 开发和使用可扩展的报告系统来评估RBP介导的拼接调制.
- 设计用于可编程拼接控制的新型工具.
主要方法:
- 开发基于 luciferase 的绑定功能拼接报告器,用于高通量选.
- 使用开发的报告员系统对718个人类RBPs的评估.
- 应用增强的交叉链接免疫沉,RNA测序和亲和力净化质谱来研究候选RBP.
- 可编程融合蛋白的工程利用已识别的外因子纳入激活域.
主要成果:
- 鉴定了718个影响替代拼接的人类RBP.
- 发现TRNAU1AP,SCAF8和RTCA的新型拼接作用,调节数百个内源拼接事件.
- 来自顶级候选RBP的强大和紧的外因子纳入激活域的表征.
- 证明工程融合蛋白在操纵外子纳入方面优于现有的人工拼接因子.
结论:
- 绑定函数测试是一种强大的工具,用于表征RBP介导的外子纳入.
- TRNAU1AP,SCAF8和RTCA是新发现的替代拼接的关键监管机构.
- 已识别的域和工程蛋白质为精确,可编程的拼接调制应用提供了有前途的新途径.
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