Also known as the ‘silencing RNA’ small-interfering RNA Interference is a double stranded RNA used to suppress the expression of genes. The gene silencing or RNA interference allows siRNA molecules to bind with RNA-induced Silencing Complex (RISC). RISC identifies the disease-causing mRNA and stops production of the protein.
Applications of Gene Silencing
Through gene silencing, RNAI offers a powerful tool to study gene expression. Expression of specific genes can be inhibited by siRNA transfection to bind the RISC complex and stop the target mRNA’s protein production. Efficient delivery of the siRNA into the cells has always been the challenge. It is equally important to consider the right cell type. There isn’t one transfection agent that works on all cells – transfection agents as well as siRNA transfection protocol are designed for particular tumor cells or primary cells. Without optimizing transfection reagents, standard transfection protocols could result in poor phenotypic response, increased toxicity, or sometimes, both.
When you’re choosing your transfection reagent, make sure you have identified a few things – which type and species of cell would you use, what is the downstream assay like and will it be a con-transfection, and are you ready to do some legwork, among other considerations. It would be great to find some literature and talk to other researchers. Someone is bound to offer their experience on something similar. Try and get a few samples from your vendor to test out, before your actual purchase. While testing you want to know if the transfection reagent will permit the siRNA to enter the cell, and if it’s without much negative effects. No to either of these questions implies you need to find another reagent.
After choosing the transfection reagent, you would want to set the siRNA transfection protocol for that particular cell. The protocol depends on the type of cell, how fast it grows, what is its density and how long it can survive, particularly after the RNA interference. The siRNA transfection protocol must also consider the method being opted for transfection, the quantity /quality of siRNA to be used as well as the time of the transfection.
siRNA Transfection in Neurons
Many studies have advocated for the application of RNAi in cultures of primary cells as well as non-neuronal cell lines. Yet, using gene silencing in neurons is difficult due to various technical as well as biological constraints. Neurons are known to show the most resistance to RNAi. A siRNA-induced delivery mechanism is only sufficient for a transient alteration of the gene’s expression, not for long-term.
Transfection of siRNA is believed to be a better delivery approach to the previous lipid nanoparticle (LNP) method. It shows similar performance across all genes and tends to have a higher response index. Unlike traditional medicine, instead of targeting only a rogue protein, siRNA transfection and RNAi attacks the problem right at the source
Advirna offers all kinds of innovative solutions in the field of molecular biology. From validated sdRNAs, custom sdRNAs and RNAi screens to starter kits, In Vivo siRNA and transfection controls, Advirna has got it all. RNAi therapeutics is changing how diseases are treated, and Advirna leads the way.