Application Note: Cell Sorting Based on RNA Detection in Living Cells Using SmartFlare™ RNA Detection Reagents
17 June 2013
Cell sorting enables the isolation of highly pure cell subpopulations, increasing the statistical significance of observed relationships between gene expression and phenotype, especially for rare events. Live cell sorting has traditionally been accomplished by detecting the presence of unique cell surface proteins, which have been identified through the use of fluorescently labeled antibodies. However, live cells cannot be sorted based on endogenous intracellular protein markers, because cells have to be fixed and permeabilized for antibody staining. Sometimes cells can be sorted on the basis of transfected reporter constructs; however, this treatment also compromises cell integrity and may perturb cellular pathways, confounding downstream analyses. In this application note read how live cells can be sorted based on RNA expression levels. The ability to detect and separate live cells based on the level of a specific RNA target provides a new opportunity to study cellular functions and identify rare cell types such as certain tumor cells and cancer stem cells.