Mechanical Fractionation of Cultured Neuronal Cells into Cell Body and Neurite Fractions.

Autor:	Arora A; Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO, USA., Goering R; Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.; RNA Bioscience Initiative, University of Colorado Anschutz Medical Campus, Aurora, CO, USA., Lo HG; Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO, USA., Taliaferro MJ; Department of Biochemistry and Molecular Genetics, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.; RNA Bioscience Initiative, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
Jazyk:	angličtina
Zdroj:	Bio-protocol [Bio Protoc] 2021 Jun 05; Vol. 11 (11), pp. e4048. Date of Electronic Publication: 2021 Jun 05 (Print Publication: 2021).
DOI:	10.21769/BioProtoc.4048
Abstrakt:	Many cells contain spatially defined subcellular regions that perform specialized tasks enabled by localized proteins. The subcellular distribution of these localized proteins is often facilitated by the subcellular localization of the RNA molecules that encode them. A key question in the study of this process of RNA localization is the characterization of the transcripts present at a given subcellular location. Historically, experiments aimed at answering this question have centered upon microscopy-based techniques that target one or a few transcripts at a time. However, more recently, the advent of high-throughput RNA sequencing has allowed the transcriptome-wide profiling of the RNA content of subcellular fractions. Here, we present a protocol for the isolation of cell body and neurite fractions from neuronal cells using mechanical fractionation and characterization of their RNA content. Graphic abstract: Fractionation of neuronal cells and analysis of subcellular RNA contents. Competing Interests: Competing interestsThe authors declare no financial or non-financial competing interests. (Copyright © The Authors; exclusive licensee Bio-protocol LLC.)
Databáze:	MEDLINE
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