Title: Efficient combinatorial targeting of RNA transcripts in single cells with Cas13 RNA Perturb-seq
Author: Wessels, Hans-Hermann, Mndez-Mancilla, Alejandro, Hao, Yuhan, Papalexi, Efthymia, Mauck, William M., Lu, Lu, Morris, John A., Mimitou, Eleni P., Smibert, Peter, Sanjana, Neville E., Satija, Rahul
Issue&Volume: 2022-12-22
Abstract: Pooled CRISPR screens coupled with single-cell RNA-sequencing have enabled systematic interrogation of gene function and regulatory networks. Here, we introduce Cas13 RNA Perturb-seq (CaRPool-seq), which leverages the RNA-targeting CRISPR–Cas13d system and enables efficient combinatorial perturbations alongside multimodal single-cell profiling. CaRPool-seq encodes multiple perturbations on a cleavable CRISPR array that is associated with a detectable barcode sequence, allowing for the simultaneous targeting of multiple genes. We compared CaRPool-seq to existing Cas9-based methods, highlighting its unique strength to efficiently profile combinatorially perturbed cells. Finally, we apply CaRPool-seq to perform multiplexed combinatorial perturbations of myeloid differentiation regulators in an acute myeloid leukemia (AML) model system and identify extensive interactions between different chromatin regulators that can enhance or suppress AML differentiation phenotypes.
DOI: 10.1038/s41592-022-01705-x
Source: https://www.nature.com/articles/s41592-022-01705-x
Nature Methods:《自然—方法学》,创刊于2004年。隶属于施普林格·自然出版集团,最新IF:47.99
官方网址:https://www.nature.com/nmeth/
投稿链接:https://mts-nmeth.nature.com/cgi-bin/main.plex
本期文章:《自然—方法学》:Online/在线发表