STAR Protocols | VOL. 3
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Cell-specific RNA purification to study translatomes of mouse central nervous system

Publication Date Jun 1, 2022

Abstract

Cell-specific RNA sequencing has revolutionized the study of cell biology. Here, we present a protocol to assess cell-specific translatomes of genetically targeted cell types. We focus on astrocytes and describe RNA purification using RiboTag tools. Unlike single-cell RNA sequencing, this approach allows high sequencing depth to detect low expression genes, and the exploration of RNAs translated in subcellular compartments. Furthermore, it avoids underestimation of transcripts from cells susceptible to cell isolation procedures. The protocol can be applied to a variety of cell types. For complete details on the use and execution of this protocol, please refer to Chai etal. (2017), Díaz-Castro etal. (2021), Díaz-Castro etal. (2019), Srinivasan etal. (2016), and Yu etal. (2018).

Concepts

Cell-specific RNA Subcellular Compartments RNA Purification RNA Sequencing Cell Types Study Of Biology High Depth Cell Isolation Procedures Mouse Central Nervous System High Sequencing Depth

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No potential conflict of interest was reported by the authors. The conception and design of the study, acquisition of data, analysis and interpretatio...

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