TNRF experts at NCATS have in-depth experience working with siRNA and RNAi screening technology to evaluate gene expression. These scientists routinely perform the following types of activities in support of NIH investigator-driven screening projects.
Common Seed Analysis to Prioritize Hits and On- or Off-Target Effects
A common source of error in RNAi experiments is when RNAi turns off genes other than the intended targets. These “off-target effects” can be detected by comparing the experimental results of an RNAi to all other similar RNAi, enabling scientists to determine if the effect observed is specific to the gene being targeted or if it occurs with all similar RNAi.
Testing Multiple siRNAs to Increase Confidence in Genes with Consensus
Because many genes are identified in error in a typical RNAi screen, follow-up experiments using different RNAi are an important way that scientists can gain confidence in their results.
Separation of False Positives from True Positives Using C911 Controls
In addition to common seed analysis, RNAi experts at NCATS have developed a novel technology to validate identified genes: C911 controls. C911 siRNAs are modified siRNAs that retain off-target effects but eliminate the normal function of the siRNA.