清脆的
计算生物学
拼车
核糖核酸
生物
引导RNA
Cas9
基因
计算机科学
遗传学
生态学
作者
Hans‐Hermann Wessels,Alejandro Méndez‐Mancilla,Efthymia Papalexi,William M. Mauck,Lu Lu,John Morris,Eleni P. Mimitou,Peter Smibert,Neville E. Sanjana,Rahul Satija
标识
DOI:10.1101/2022.02.02.478894
摘要
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 array which 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.
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