RNA Interference (RNAi)
Research tools designed for RNA interference studies are used to suppress protein expression transiently or permanently, or to investigate the role of small RNA molecules in regulating gene expression.
The T7 RiboMAX™ Express RNAi System is an in vitro transcription system designed for efficient synthesis of milligram amounts of short interfering RNAs (siRNAs) or hairpin siRNAs (shRNAs) for mammalian RNAi studies.
The psiCHECK™ Vectors provide a quantitative and rapid approach for initial optimization of RNA interference. These vectors also are ideal for examining the effect of 3´ untranslated regions (3´ UTRs), such as miRNA target sequences, on gene expression.
The pmirGLO Vector is designed to quantitatively evaluate microRNA (miRNA) activity by the insertion of miRNA target sites downstream or 3´ of the firefly luciferase gene (luc2). Reduced firefly luciferase expression indicates the binding of endogenous or introduced miRNAs to the cloned miRNA target sequence.
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RNA Interference Basics
In RNAi, small double-stranded RNAs suppress expression of a target protein by stimulating the specific degradation of the complementary target mRNA. RNAi is a powerful technique that allows researchers to investigate gene function by knocking out or knocking down the level of a particular protein. RNAi is a multistep process in which the dsRNA is recognized by an RNase III family member (e.g., Dicer in Drosophila) and is cleaved into siRNAs of 21–23 nucleotides. The siRNAs are incorporated into an RNAi targeting complex known as RISC (RNA-induced silencing complex), which destroys mRNAs homologous to the siRNA contained in the complex, effectively knocking out protein expression.
MicroRNAs (miRNAs) are short, noncoding RNAs that regulate gene expression. miRNAs target the 3´-untranslated region of messenger RNA sequences, affect RNA stability and prevent translation. miRNAs have been implicated them in a wide range of biological processes and disease states including development, metabolism, and cancer.