Detection of siRNA-Mediated Gene Modulation Using SmartFlare™ Detection Probes
Modulation of gene expression using techniques such as RNAi has become a fundamental tool in the study of gene function and biological pathways. To determine the efficiency of gene knockdown using most widely used detection methods, cell samples must be sacrificed by lysis or permeabilization and fixation. In addition to sample destruction, another disadvantage of these techniques is that they yield results that only reflect the average expression of the gene in the collected cell population. One nondestructive option is the use of transfected reporter constructs. However, although the cells remain alive, the technique cannot reveal endogenous gene expression and can have negative effects on cell health. An ideal RNA detection agent provides a noninvasive approach to interrogating gene expression while enabling sorting of live cells that can be separated and directly used for downstream studies.
Benefits of Using SmartFlare™ RNA Detection Probes for Detecting Gene Modulation
SmartFlare™ RNA detection probes can detect target mRNA and microRNA levels in live, intact cells. SmartFlare™ probes require no carrier agent to enter cells and have no toxic effect on cell health. SmartFlare™ probes enable users to quickly verify gene expression, and, in conjunction with cell sorting, isolate desired cell populations. Furthermore, these reagents require no sample preparation and leave cells intact after the detection event, allowing for downstream studies.
Using SmartFlare™ RNA detection probes, we have demonstrated accurate, efficient detection of siRNA-mediated gene knockdown using a SmartFlare™ probe specific for a target of interest. SmartFlare™ probes enable fast and accurate detection of target gene expression at single cell resolution. The ability to specifically detect RNA levels on a cell–by-cell basis provides new opportunities to link biological pathways and physiological processes to gene functions.