In Vivo siRNA Transfection (animal siRNA delivery)
In vivo methods require live animals systems where the desired molecule is delivered into the animal or locally to specific tissues. The immune response of the animal will influence the transfection efficiency. By targeting tissue types, such as the brain versus the kidney, a researcher can study the organ’s specific effects during in vivo transfection. Though complications do arise, for example nucleic acids sensitive to in vivo degradation, such as siRNAs and microRNAs are degraded by RNases in vivo within seconds after administration, using liposome encapsulation, condensing to the nanoparticle size, and using optimized in vivo transfection protocol and tissue-targeted reagents (see In Vivo Nanoparticle Transfection Kit) can significantly improve and prolong stability in vivo.
In animals and plants, RNA interference is part of an immune response to viruses or injection of foreign genetic materials into the cellular structure. This process is recreated by scientists to study gene function through RNAi gene silencing; commercially synthesized siRNAs can be ordered and utilized as a key part of completely separate research on account of siRNA’s crucial role in many experimental processes.
Because of the robust and selective effect RNAi has on gene expression, it has become an invaluable research tool for studies including in vivo target validation studies using animal models. The major challenge in performing RNAi studies in vivo is the effective, directed delivery of functional siRNA, shRNA, and miRNA molecules into specific tissues. Altogen® In Vivo Transfection Reagents can be conjugated with siRNA and administered intratumorally (i.t) or systemically via an intravenous (i.v) tail vein injection in order to provide directed gene silencing in specific tissues, including liver, pancreas, kidney, and tumors, and selective knockdown can be seen as early as 24 hours after injection.
Gene silencing by RNA Interference (RNAi) is a powerful research tool for studying gene function in mammalian cells. RNAi is a biological phenomenon by which double stranded RNA (dsRNA) specifically reduces gene expression of its corresponding gene. Potent inhibition of specific gene expression is experimentally achieved by the transfection of small interfering RNA (siRNA), which completes the RISC complex and causes degradation of target complementary mRNA molecules in the cell. Therefore, successful, potent RNAi experimentation is dependent upon the efficient delivery of the siRNA into cells via transfection of stable and functional siRNA molecules. There is a plethora of protocols and methods that require careful selection to maximize the success of this process.
Most DNA transfection reagents are incompatible with siRNA (which is sensitive to serum) or are capable of inducing cytotoxicity, which negatively affects gene expression studies. Altogen® Transfection Reagents have been developed and optimized specifically for use with siRNA transfection and allow transfection to take place in the presence of serum without lowering the transfection efficiency or reducing cell viability. Ready-to-run transfection protocols eliminate the need for extensive siRNA optimization experiments. Altogen Biosystems kits are compatible with transfection of any negatively charged molecules (including protein, RNA, DNA, and small molecule compounds).
Featured in vivo transfection products from Altogen Biosystems:
Altogen Custom Services provides in vivo transfection services and other specialized biotechnology and pharmaceutical services, our contract research services (CRO) are GLP-compliant and include over 50 validated xenograft models, development of stable cell lines, RNA Interference (RNAi) services, assay development, ELISA and Western Blot services, siRNA library screening and transfection services. Generation of stably-expressing cell lines can be very expensive and time-consuming. Altogen Labs offers generation of stable cell line service completed in just 28 days (see service details).