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科学家开发出一种用于大规模体内CRISPR筛选的平台
作者:小柯机器人 发布时间:2021/8/22 14:10:42

美国犹他大学Randall T. Peterson课题组开发出一种用于大规模体内CRISPR筛选的平台。相关论文于2021年8月19日在线发表在《科学》杂志上。

研究人员报告了Multiplexed Intermixed CRISPR Droplets (MIC-Drop),这是一个结合了液滴微流控技术、单针大规模CRISPR核糖核酸蛋白注射和DNA条形码的平台,能够实现斑马鱼的大规模功能基因筛选。该平台可以有效地鉴定负责形态学或行为表型的基因。在一个应用中,研究人员显示MIC-Drop可以识别小分子靶点。此外,在对188个表征不明确的基因进行的MIC-Drop筛选中,研究人员发现了几个对心脏发育和功能很重要的基因。由于MIC-Drop有可能扩展到数千个基因,它可以在模式生物中进行基因组规模的反向遗传筛选。

据了解,CRISPR-Cas9可以被放大用于培养细胞的大规模筛选,但在动物中进行CRISPR筛选一直是个挑战,因为生成、验证和跟踪大量的突变动物是很困难的。

附:英文原文

Title: MIC-Drop: A platform for large-scale in vivo CRISPR screens

Author: Saba Parvez, Chelsea Herdman, Manu Beerens, Korak Chakraborti, Zachary P. Harmer, Jing-Ruey J. Yeh, Calum A. MacRae, H. Joseph Yost, Randall T. Peterson

Issue&Volume: 2021/08/19

Abstract: CRISPR-Cas9 can be scaled up for large-scale screens in cultured cells, but CRISPR screens in animals have been challenging because generating, validating, and keeping track of large numbers of mutant animals is prohibitive. Here, we report Multiplexed Intermixed CRISPR Droplets (MIC-Drop), a platform combining droplet microfluidics, single-needle en masse CRISPR ribonucleoprotein injections, and DNA barcoding to enable large-scale functional genetic screens in zebrafish. The platform can efficiently identify genes responsible for morphological or behavioral phenotypes. In one application, we show MIC-Drop can identify small molecule targets. Furthermore, in a MIC-Drop screen of 188 poorly characterized genes, we discover several genes important for cardiac development and function. With the potential to scale to thousands of genes, MIC-Drop enables genome-scale reverse-genetic screens in model organisms.

DOI: 10.1126/science.abi8870

Source: https://science.sciencemag.org/content/early/2021/08/18/science.abi8870

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:41.037