使用CRISPR-Cas9基因编辑标记内源蛋白质的HiBiT协议.
Kaylee P Lankford1, John D Hulleman2
1Department of Ophthalmology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390, USA.
STAR protocols
|April 10, 2024
概括
这项研究引入了一种结合CRISPR-Cas9基因组编辑和NanoBiT系统的新方法,用于高效的体外和体内蛋白质检测. 这种方法简化了内源性蛋白质的高通量选.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 生物技术是生物技术.
背景情况:
- 精确的蛋白质检测对于理解细胞功能至关重要.
- 对于内源蛋白质分析的现有方法可能是复杂的和低通量.
研究的目的:
- 介绍一种新的,简化的 in vitro 和 in vivo 蛋白质检测方法.
- 通过使用CRISPR-Cas9和NanoBiT,使内源蛋白质的高通量选成为可能.
主要方法:
- 整合CRISPR-Cas9基因组编辑与NanoBiT蛋白质检测系统.
- 细胞培养,核糖蛋白输送和细胞监测的详细协议.
- 使用HiBiT测定,基因组DNA分析和HiBiT涂抹的编辑分析.
主要成果:
- 成功实施了一种简单且多功能蛋白质检测协议.
- 通过各种分析技术展示编辑特异性.
- 促进内源性蛋白质的高通量选能力.
结论:
- 提出的方法为蛋白质检测提供了一个简单,高效和多功能工具.
- 这种方法显著提高了对内源蛋白进行高通量选的易度.
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