Protocol to identify genes required for cardiomyocyte development using Perturb-Seq
Year: 2026
Type: article
Abstract: While Perturb-Seq combines CRISPR-based screening with single-cell RNA sequencing (scRNA-seq), large-scale experiments are costly and its application during development is complicated by differentiation heterogeneity. Here, we present a protocol to identify genes required for cardiomyocyte development using Perturb-Seq. We describe steps for sgRNA (single guide RNA) library cloning and infection, cardiomyocyte differentiation, cell hashing, super loading, and scRNA-seq. We then detail procedures for sequencing, mapping, and data analysis. For complete details on the use and execution of this protocol, please refer to Sivakumar et al. 1
Source: STAR Protocols
Institution: The University of Texas Southwestern Medical Center
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
Funders: U.S. Department of Defense, Burroughs Wellcome Fund, Cancer Prevention and Research Institute of Texas, University of Texas Southwestern Medical Center, National Institutes of Health, National Heart, Lung, and Blood Institute, National Human Genome Research Institute, National Institute of General Medical Sciences
Awards: Systematic Assessment of Combinatorial Transcription Factor Activity, G390011392, Comprehensive Evaluation of Functional Enhancers in Breast Cancer Risk Susceptibility Loci, Combinatorial Biology of Gene Regulation for Cellular Engineering, G6245228462, Defining regenerative potential in the cardiac conduction system, Multiscale functional characterization of genomic variation in human developmental disorders, Exploring Mechanisms of Cardiac Pacemaker Cell Fate Determination
OpenAlex ID: W7204200149