Difficulties of Transfection
Transfection as a technique is one of the most versatile tools available to molecular biologists and geneticists. Although it is superficially simple and easy to understand, the details of performing it are a bit more complicated; finding the optimal conditions for cellular uptake of genetic material and designing cargo vesicles is difficult. Thankfully many companies now offer kits for transfecting specific cell lines with specific kinds of nucleic acids.
One of the most important aspects of transfection is the kind of nucleic acid being transfected – be it short siRNA or large plasmids encoding full proteins, transfection will work to bring them into a cell. However, each individual kind of nucleic acid differs in the optimal conditions for its function. siRNA, for example, is notorious for being difficult to transfect because it is not particularly stable in most conventional buffers. With these challenges in mind, several companies have set off to improve the assortment of tools available to labs for their transfection needs.
Corporate Products and Services
The technical difficulties associated with inefficient DNA, siRNA, or miRNA delivery into cells can lead to time consuming trials that would otherwise be avoided with pre-optimized products. Some challenges include the limited stability of nucleic acids, inefficient transfer of material across the cell membrane, and poor PK/PD efficacy in vivo. Stable and functional DNA or RNA molecules typically require chemical modifications and carrier molecule encapsulation (liposome-, lipid-, polymer-, etc.) in order to ensure efficient functional effects and minimized off-target effects.
Many companies provide both pre-optimized reagents and advice to laboratories conducting their own transfection experiments. Transfection kits can save plenty of time in short-run experiments, while corporate advice can help in larger-scale experiments involving high-throughput screenings of cells. As an important example, siRNA transfection has been used to determine the broad effects of silencing certain genes in cells, but the process of screening is difficult and time consuming. It may be worth the resources to outsource a large-scale screen to a company with experience in the field.
Expert advice is also important for designing specific transfection experiments that require either stably-transfected cells or specific attention to off-target effects. Altogen Services offers contract research services to pharmaceutical, biotech and academic institutions worldwide, with a large number of offerings of custom research services. One of the products that Altogen offers is a Nanoparticle In Vivo Reagent. This reagent is administered intratumorally via injection or systemic intravenous injection. The nanoparticle complexes containing a siRNA or plasmid DNA cargo are stable in a serum for 16 hours and can be delivered efficiently to a number of tissues and tumor types. Other in vivo delivery products include lipid-, polymer-, and PEG-Liposome based reagents that are used to protect siRNA from degradation and effectively deliver cargo to the targeted tissue or tumor. An example of a cell line specific transfection reagent is the A549 Transfection Reagent that provides highly efficient delivery of miRNA, siRNA and plasmid DNA into A549 cells with low cytotoxicity.
Altogen Biosystems is a biotechnology company developing cell line specific, pre-optimized transfection reagents for in vitro and in vivo delivery of biomolecules. Molecular Biology labs typically use transfections in order to study gene function and regulation, as well as to perform protein production, biochemical analysis, mutational analysis and gene expression modulation.