小柯机器人

美国纽约大学Neville E. Sanjana研究组利用可扩展的单细胞CRISPR筛选解析染色质可及性的遗传决定因素。该研究于2021年4月29日在线发表于国际一流学术期刊《自然—生物技术》。

研究人员报道了一种用于转座酶可及性染色质的CRISPR单细胞组合索引测定法（CRISPR–sciATAC），可将遗传扰动与大量细胞中全基因组的染色质可及性联系起来。在人类骨髓性白血病细胞中，研究人员将CRISPR–sciATAC应用于105个染色质相关基因，从而产生约30,000单细胞的染色质可及性数据。研究人员将特定染色质重塑因子的丢失与全局可及性以及单个转录因子（TFs）结合位点的变化联系起来。

例如，研究人员表明H3K27甲基转移酶EZH2的丢失会增加参与胚胎发育异染色区的可及性，并触发HOXA和HOXD簇中基因的表达。在调控位点的亚群中，研究人员还分析了染色质重塑因子丢失后核小体间距的变化。CRISPR-sciATAC是一种高通量单细胞方法，可用于研究正常状态和疾病状态下遗传扰动对染色质的影响。

据悉，CRISPR筛选已用于将遗传扰动与基因表达和表型的变化联系起来。

附：英文原文

Title: Profiling the genetic determinants of chromatin accessibility with scalable single-cell CRISPR screens

Author: Noa Liscovitch-Brauer, Antonino Montalbano, Jiale Deng, Alejandro Mndez-Mancilla, Hans-Hermann Wessels, Nicholas G. Moss, Chia-Yu Kung, Akash Sookdeo, Xinyi Guo, Evan Geller, Suma Jaini, Peter Smibert, Neville E. Sanjana

Issue&Volume: 2021-04-29

Abstract: CRISPR screens have been used to connect genetic perturbations with changes in gene expression and phenotypes. Here we describe a CRISPR-based, single-cell combinatorial indexing assay for transposase-accessible chromatin (CRISPR–sciATAC) to link genetic perturbations to genome-wide chromatin accessibility in a large number of cells. In human myelogenous leukemia cells, we apply CRISPR–sciATAC to target 105chromatin-related genes, generating chromatin accessibility data for ~30,000single cells. We correlate the loss of specific chromatin remodelers with changes in accessibility globally and at the binding sites of individual transcription factors (TFs). For example, we show that loss of the H3K27 methyltransferase EZH2 increases accessibility at heterochromatic regions involved in embryonic development and triggers expression of genes in the HOXA and HOXD clusters. At a subset of regulatory sites, we also analyze changes in nucleosome spacing following the loss of chromatin remodelers. CRISPR–sciATAC is a high-throughput, single-cell method for studying the effect of genetic perturbations on chromatin in normal and disease states.

DOI: 10.1038/s41587-021-00902-x

Source: https://www.nature.com/articles/s41587-021-00902-x

Nature Biotechnology：《自然—生物技术》，创刊于1996年。隶属于施普林格·自然出版集团，最新IF：68.164
官方网址：https://www.nature.com/nbt/
投稿链接：https://mts-nbt.nature.com/cgi-bin/main.plex