小柯机器人

美国纽约大学Rahul Satija等研究人员合作利用Cas13 RNA Perturb-seq对单细胞中的RNA转录本进行有效的组合定位。该研究于2022年12月22日在线发表于国际一流学术期刊《自然—方法学》。

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