Electroporation in a Pipette Tip
Christofer Cunning, Ph.D., senior marketing manager at Life Technologies, says that reproducibility, toxicity concerns, and the ability to successfully transfect primary cells and difficult-to-transfect cells, including immune cells and stem cells, remain challenging in siRNA experiments. “In general, people have used vector approaches with viral delivery systems for difficult transfections,” he says. “Use of viral vectors, however, remains time-consuming, requires cloning steps, is relatively expensive initially, and may require significant safety precautions.” In addition, while electroporation has been widely applied, “it requires strict adherence to already set protocols and a lot of siRNA.”
To address these issues, Invitrogen launched its Neon™ transfection system earlier this year. In contrast to other systems, Neon allows transfection to take place in a pipette tip, using as few as 2x104 cells. “This technology is amenable to changes in protocols that can be defined by the user,” Dr. Cunning asserts.
Unlike standard cuvette-based electroporation chambers, the Neon system uses a biologically compatible pipette tip chamber that generates a more uniform electric field. The company says this design allows better maintenance of physiological conditions resulting in high cell survival compared to conventional electroporation. A universal reagent kit allows use with all cell types.
Invitrogen recently launched Invivofectamine™ for in vivo siRNA transfection. The company claims that in contrast to other marketed products, Invivofectamine can be injected in small volumes (microliters as opposed to milliliters) and without high pressure, thereby minimizing the potential of inconsistent results and gross harm to the subject animal.
This reagent also reportedly provides stability to siRNA so that it remains intact and ready to perform the selected knockdown. Invitrogen has demonstrated successful delivery of intact siRNA molecules via direct injection into established tumors, tail vein injection to the liver, kidney, lungs, spleen, and pancreas, as well as effective and specific knockdown in these tissues.