Perturb-tracing enables high-content screening of multiscale 3D genome regulators

作者：Yubao Cheng, Mengwei Hu, Bing Yang, Tyler B Jensen, Tianqi Yang, Ruihuan Yu, Zhaoxia Ma, Jonathan S. D. Radda, Shengyan Jin, Chongzhi Zang, Siyuan Wang · 发表于：bioRxiv (Cold Spring Harbor Laboratory) · 年份：2023 · DOI：10.1101/2023.01.31.525983 · 被引用次数：12 · 研究领域：Cell Image Analysis Techniques、Single-cell and spatial transcriptomics、Genomics and Chromatin Dynamics

Abstract Three-dimensional (3D) genome organization becomes altered during development, aging, and disease 1–23 , but the factors regulating chromatin topology are incompletely understood and currently no technology can efficiently screen for new regulators of multiscale chromatin organization. Here, we developed an image-based high-content screening platform (Perturb-tracing) that combines pooled CRISPR screen, a new cellular barcode readout method (BARC-FISH), and chromatin tracing. We performed a loss-of-function screen in human cells, and visualized alterations to their genome organization from 13,000 imaging target-perturbation combinations, alongside perturbation-paired barcode readout in the same single cells. Using 1.4 million 3D positions along chromosome traces, we discovered tens of new regulators of chromatin folding at different length scales, ranging from chromatin domains and compartments to chromosome territory. A subset of the regulators exhibited 3D genome effects associated with loop-extrusion and A-B compartmentalization mechanisms, while others were largely unrelated to these known 3D genome mechanisms. We found that the ATP-dependent helicase CHD7, the loss of which causes the congenital neural crest syndrome CHARGE 24 and a chromatin remodeler previously shown to promote local chromatin openness 25–27 , counter-intuitively compacts chromatin over long range in different genomic contexts and cell backgrounds including neural crest cells, and globally rep...