Probe-Seq: Method for RNA Sequencing of Specific Cell Types from Animal Tissue

Citation:

Amamoto R, Garcia MD, West ER, Choi J, Lapan SW, Lane EA, et al. Probe-Seq: Method for RNA Sequencing of Specific Cell Types from Animal Tissue. Bio Protoc. 2020;10 (18) :e3749.

Date Published:

2020 Sep 20

Abstract:

Most organs and tissues are composed of many types of cells. To characterize cellular state, various transcription profiling approaches are currently available, including whole-tissue bulk RNA sequencing, single cell RNA sequencing (scRNA-Seq), and cell type-specific RNA sequencing. What is missing in this repertoire is a simple, versatile method for bulk transcriptional profiling of cell types for which cell type-specific genetic markers or antibodies are not readily available. We therefore developed Probe-Seq, which uses hybridization of gene-specific probes to RNA markers for isolation of specific types of cells, to enable downstream FACS isolation and bulk RNA sequencing. We show that this method can enable isolation and profiling of specific cell types from mouse retina, frozen human retina, midgut, and developing chick retina, suggesting that it is likely useful for most organisms.