siRNA Transfection

Gene silencing or knockdown can be done easily now in most cell types, but there are a few things you need to consider before starting:

• First you’ll need to design the siRNA (small interferring RNA) against your gene
• Second you’ll need to deliver it to the cells effectively with a siRNA transfection procedure

siRNAs are easily introduced into cells with a siRNA transfection reagent . Soon after being inserted in the mammalian cell, the siRNA molecules become a part of the RNA-induced silencing complex (RISC). Guided by the antisense strand of the siRNA, RISC degrades the targeted mRNA inhibiting its translation. Assays are then performed to detect the RNAi activity. Controls are normally set up so RNAi results can be properly compared.

The success of RNAi is dependant on correct delivery of siRNA in appropriate amount at a time when it will brings about the maximum expected response. Such precision can be tricky. Off-targeting by siRNAs proves lethal and poses analytical issues at times. Researchers are looking for better ways of designing and delivering siRNA.