Department of Dermatology, University of California San Diego, La Jolla, California, USA.
Department of Dermatology, University of California San Diego, La Jolla, California, USA.
J Invest Dermatol. 2020 Apr;140(4):723-728.e1. doi: 10.1016/j.jid.2020.01.018.
CRISPR and Cas proteins, often referred to as CRISPR/Cas, are the components of a bacterial genome editing system that can be used to perturb genes in cells and tissues. A classic application is to use CRISPR/Cas to generate genetic loss-of-function. When performed at large scale and combined with deep sequencing techniques, CRISPR-based perturbations can be performed in a high throughput setting to screen many candidate genomic elements for their roles in a phenotype of interest. Here, we discuss major considerations in the design, execution, and analysis of CRISPR screens. We focus on CRISPR knockout screens but also review adaptations to the CRISPR/Cas system that highlight the versatility of the system to make other types of experimental genetic changes as well. We also discuss examples of CRISPR genetic screens in investigative dermatology and how they may be used to answer key scientific questions in the field.
CRISPR 和 Cas 蛋白,通常被称为 CRISPR/Cas,是细菌基因组编辑系统的组成部分,可以用于干扰细胞和组织中的基因。一个经典的应用是使用 CRISPR/Cas 来产生基因功能丧失。当大规模进行并结合深度测序技术时,基于 CRISPR 的干扰可以在高通量设置中进行,以筛选许多候选基因组元件在感兴趣的表型中的作用。在这里,我们讨论了 CRISPR 筛选设计、执行和分析中的主要考虑因素。我们专注于 CRISPR 敲除筛选,但也回顾了对 CRISPR/Cas 系统的改编,这些改编突出了该系统的多功能性,使其能够进行其他类型的实验性遗传改变。我们还讨论了 CRISPR 遗传筛选在调查性皮肤病学中的应用,以及它们如何用于回答该领域的关键科学问题。