Department of Animal Center, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Adv Exp Med Biol. 2018;1068:171-176. doi: 10.1007/978-981-13-0502-3_14.
Functional genomics aims to develop an in-depth understanding of how specific gene dysfunctions are related to diseases. A common method for investigating the genome and its complex functions is via perturbation of the interactions between the DNA, RNA and their protein respective protein derivatives. Commonly, arrayed and pooled genetic screens are utilized to achieve this and in recent years have been fundamental in achieving the current level of understanding for gene dysfunctions. However, they are limited in specific aspects which scientists have attempted to address. Clustered regularly palindromic repeats (CRISPR)-based methods for genetic screens have in recent years become more prevalent but crucially shared similar properties to previous methods and failing to provide a distinct advantage over previous methods. CROP-seq, Perturb-seq, and CRISPR-seq have combined CRISPR and single-cell RNA-sequencing (scRNA-seq) and is the newest addition to the geneticist's arsenal, providing scientists with methods to edit DNA with improved speed, accuracy, and efficiency which could usher us into a new era of study methods for functional genomics. We briefly overview the CRISPR-Cas9 systems, the evolution of genetic screening in recent years, and evaluate and discuss the significance of CROP-seq, Perturb-seq, and CRISPR-seq.
功能基因组学旨在深入了解特定基因功能障碍与疾病的关系。研究基因组及其复杂功能的常用方法是通过干扰 DNA、RNA 及其相应蛋白质衍生物之间的相互作用来实现。通常，通过排列和汇集遗传筛选来实现这一点，近年来在实现对基因功能障碍的当前理解水平方面发挥了重要作用。然而，它们在某些方面存在局限性，科学家们一直在试图解决这些问题。近年来，基于成簇规律间隔短回文重复（CRISPR）的遗传筛选方法越来越流行，但关键是它们与以前的方法具有相似的特性，并没有提供明显优于以前方法的优势。CROP-seq、Perturb-seq 和 CRISPR-seq 结合了 CRISPR 和单细胞 RNA 测序（scRNA-seq），是遗传学家武器库中的最新成员，为科学家提供了具有更高速度、准确性和效率的编辑 DNA 的方法，这可能将我们带入功能基因组学研究方法的新时代。我们简要概述了 CRISPR-Cas9 系统，以及近年来遗传筛选的发展，并评估和讨论了 CROP-seq、Perturb-seq 和 CRISPR-seq 的意义。
