Transcriptional profiling of cells sorted by RNA abundance

Nat Methods. 2014 May;11(5):549-551. doi: 10.1038/nmeth.2910. Epub 2014 Mar 30.

Abstract

We have developed a quantitative technique for sorting cells on the basis of endogenous RNA abundance, with a molecular resolution of 10-20 transcripts. We demonstrate efficient and unbiased RNA extraction from transcriptionally sorted cells and report a high-fidelity transcriptome measurement of mouse induced pluripotent stem cells (iPSCs) isolated from a heterogeneous reprogramming culture. This method is broadly applicable to profiling transcriptionally distinct cellular states without requiring antibodies or transgenic fluorescent proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Alleles
  • Animals
  • Cell Culture Techniques*
  • Cellular Reprogramming
  • Doxycycline / chemistry
  • Embryonic Stem Cells / cytology
  • Fibroblasts / metabolism
  • Flow Cytometry
  • Gene Expression Profiling*
  • Genome-Wide Association Study
  • Green Fluorescent Proteins / metabolism
  • In Situ Hybridization, Fluorescence
  • Induced Pluripotent Stem Cells / cytology*
  • Mice
  • NIH 3T3 Cells
  • Oligonucleotide Array Sequence Analysis
  • Polymerase Chain Reaction
  • RNA / metabolism*
  • RNA, Messenger / metabolism
  • Transcription, Genetic*
  • Transgenes

Substances

  • RNA, Messenger
  • Green Fluorescent Proteins
  • RNA
  • Doxycycline

Associated data

  • GEO/GSE55671
  • GEO/GSE55672
  • GEO/GSE55919