美国斯坦福大学Howard Y. Chang小组发现，双向表观遗传编辑揭示基因调控的层次结构。该项研究成果于2024年5月17日在线发表在《自然—生物技术》杂志上。

据悉，CRISPR扰动方法在研究非编码元件和基因相互作用方面能力有限。

附：英文原文

Title: Bidirectional epigenetic editing reveals hierarchies in gene regulation

Author: Pacalin, Naomi M., Steinhart, Zachary, Shi, Quanming, Belk, Julia A., Dorovskyi, Dmytro, Kraft, Katerina, Parker, Kevin R., Shy, Brian R., Marson, Alexander, Chang, Howard Y.

Issue&Volume: 2024-05-17

Abstract: CRISPR perturbation methods are limited in their ability to study non-coding elements and genetic interactions. In this study, we developed a system for bidirectional epigenetic editing, called CRISPRai, in which we apply activating (CRISPRa) and repressive (CRISPRi) perturbations to two loci simultaneously in the same cell. We developed CRISPRai Perturb-seq by coupling dual perturbation gRNA detection with single-cell RNA sequencing, enabling study of pooled perturbations in a mixed single-cell population. We applied this platform to study the genetic interaction between two hematopoietic lineage transcription factors, SPI1 and GATA1, and discovered novel characteristics of their co-regulation on downstream target genes, including differences in SPI1 and GATA1 occupancy at genes that are regulated through different modes. We also studied the regulatory landscape of IL2 (interleukin-2) in Jurkat T cells, primary T cells and chimeric antigen receptor (CAR) T cells and elucidated mechanisms of enhancer-mediated IL2 gene regulation. CRISPRai facilitates investigation of context-specific genetic interactions, provides new insights into gene regulation and will enable exploration of non-coding disease-associated variants.

DOI: 10.1038/s41587-024-02213-3

Source: https://www.nature.com/articles/s41587-024-02213-3