Detection of low abundance RNA molecules in individual cells by flow cytometry

A variety of RNA analysis technologies are available for the detection of RNA transcripts from bulk cell populations. However, the techniques for RNA detection from individual cells have been limited. Here we adapt a novel in situ signal amplification method (the RNAScope® detection platform) for th...

Full description

Saved in:

Bibliographic Details
Published in:	PloS one 2013-02, Vol.8 (2), p.e57002-e57002
Main Authors:	Hanley, Mary Beth, Lomas, Woodrow, Mittar, Dev, Maino, Vernon, Park, Emily
Format:	Article
Language:	eng
Subjects:	Abundance BCR protein Biology Blood & organ donations Cell Line Correlation analysis Cytometry Deoxyribonucleic acid DNA DNA probes Flow cytometry Flow Cytometry - methods Gene expression HIV Core Protein p24 - genetics HIV Infections HIV testing Humans Hybridization Infections Leukocytes, Mononuclear - metabolism Leukocytes, Mononuclear - virology Messenger RNA Nucleic Acid Hybridization R&D Reproducibility of Results Research & development Ribonucleic acid RNA RNA - chemistry RNA - genetics RNA, Messenger - chemistry RNA, Messenger - genetics Sensitivity analysis Sensitivity and Specificity Target detection Telomerase Viral infections
Online Access:	Get full text
Tags:	Add Tag No Tags, Be the first to tag this record!

Description
Summary:	A variety of RNA analysis technologies are available for the detection of RNA transcripts from bulk cell populations. However, the techniques for RNA detection from individual cells have been limited. Here we adapt a novel in situ signal amplification method (the RNAScope® detection platform) for the analysis of intracellular RNAs in individual cells by flow cytometry. Using novel target-specific probes that were designed to suppress background signals, we demonstrate the specific detection of HIV gag RNAs in HIV-infected cellular samples, in addition to bcr and abl mRNAs in the K562 cell line. This method was capable of distinguishing cells expressing low abundance RNA transcripts and correlated well with quantitative imaging analysis. Furthermore, multiple distinct RNA targets were simultaneously detected with a high specificity without interference. Overall, the sensitivity and specificity of this method will be useful for the analysis of functionally important RNA species from individual cells, even at very low copy numbers.
ISSN:	1932-6203 1932-6203