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Protocol for Single-Nucleus Transcriptomics of Diploid and Tetraploid Cardiomyocytes in Murine Hearts

Miao Cui, Eric N. Olson

2020STAR Protocols25 citationsDOIOpen Access PDF

Abstract

Murine cardiomyocytes undergo proliferation, multinucleation, and polyploidization during the first 3 weeks of postnatal life, resulting in a mixture of diploid and tetraploid cardiomyocytes in the heart. Understanding the molecular differences between diploid and tetraploid cardiomyocytes from these processes has been limited due to lack of unique markers and their heterogenous origins. Here, we apply single-nucleus RNA-sequencing to fluorescence-activated cell sorting-selected diploid and tetraploid cardiomyocytes to characterize their heterogeneity and molecular distinctions. For complete details on the use and execution of this protocol, please refer to Cui et al. (2020).

Topics & Concepts

PloidyCell sortingNucleusBiologyTranscriptomeRNACell biologySortingCellGeneticsGeneGene expressionComputer scienceProgramming languageCongenital heart defects researchSingle-cell and spatial transcriptomicsCardiomyopathy and Myosin Studies
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