Your browser doesn't support javascript.
loading
Probe-Seq: Method for RNA Sequencing of Specific Cell Types from Animal Tissue.
Amamoto, Ryoji; Garcia, Mauricio D; West, Emma R; Choi, Jiho; Lapan, Sylvain W; Lane, Elizabeth A; Perrimon, Norbert; Cepko, Constance L.
Afiliação
  • Amamoto R; Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Blavatnik Institute, Harvard Medical School, Boston MA 02115, USA.
  • Garcia MD; Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Blavatnik Institute, Harvard Medical School, Boston MA 02115, USA.
  • West ER; Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Blavatnik Institute, Harvard Medical School, Boston MA 02115, USA.
  • Choi J; Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Blavatnik Institute, Harvard Medical School, Boston MA 02115, USA.
  • Lapan SW; Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Blavatnik Institute, Harvard Medical School, Boston MA 02115, USA.
  • Lane EA; Department of Genetics, Howard Hughes Medical Institute, Blavatnik Institute, Harvard Medical School, Boston MA 02115, USA.
  • Perrimon N; Department of Genetics, Howard Hughes Medical Institute, Blavatnik Institute, Harvard Medical School, Boston MA 02115, USA.
  • Cepko CL; Departments of Genetics and Ophthalmology, Howard Hughes Medical Institute, Blavatnik Institute, Harvard Medical School, Boston MA 02115, USA.
Bio Protoc ; 10(18): e3749, 2020 Sep 20.
Article em En | MEDLINE | ID: mdl-33659409
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, Drosophila midgut, and developing chick retina, suggesting that it is likely useful for most organisms.
Palavras-chave

Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2020 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2020 Tipo de documento: Article