Quantitative PCR primer design affects quantification of dsRNA-mediated gene knockdown.

ABSTRACT: RNA interference (RNAi) is a powerful tool for studying functions of candidate genes in both model and nonmodel organisms and a promising technique for therapeutic applications. Successful application of this technique relies on the accuracy and reliability of methods used to quantify gene knockdown. With the limitation in the availability of antibodies for detecting proteins, quantitative PCR (qPCR) remains the preferred method for quantifying target gene knockdown after dsRNA treatment. We evaluated how qPCR primer binding site and target gene expression levels affect quantification of intact mRNA transcripts following dsRNA-mediated RNAi. The use of primer pairs targeting the mRNA sequence within the dsRNA target region failed to reveal a significant decrease in target mRNA transcripts

SUBMITTER: Onchuru TO 

PROVIDER: S-EPMC6662389 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature