Category: Cellular Technologies
Cellular transfection methods have been relied upon for several decades in the fields of gene therapy, drug discovery, compound screening, and basic biology research. However, traditional biological, chemical, and physical transfection methods have challenges, including low transfection efficiency rates, high cell toxicity, and inadequate scalability. Here we describe a novel method to introduce genetic material and proteins into mammalian cells, using ultrasonic irradiation of encapsulated microbubbles in a process called sonoporation.
Microbubbles are functionalized with an antibody, allowing researchers to target their cells of choice. Next, we combined the transfection material, which may be nucleic acids, proteins, or small molecules, with the functionalized microbubbles. The antibody coated microbubbles bind to the cells during incubation. The cells are then irradiated with ultrasonic energy, exciting oscillations and deformations of the microbubbles that in turn create transient pores in the cell membrane. This results in enhanced delivery of the desired material into the cells.
We demonstrate proof of concept of this novel transfection method using both plasmid DNA and ribonucleoprotein complexes (RNPs), loaded onto streptavidin-coated microbubbles functionalized with biotinylated anti-CD51 antibody. Plasmid DNA transfects at low efficiencies with almost no cell death, as measured by FACS. CRISPR RNP complexes are transfected at higher efficiencies and maintain their ability to edit the genome. The low cell death rate is advantageous for cell lines that are normally difficult to transfect. Sonoporation enables researchers who work with cells such as lymphocytes or stem cells an opportunity to pursue plasmid screening, CRISPR editing, gene expression profiling, single-cell sequencing, and drug screening and delivery in high throughput workflows.
Marsha Blauwkamp– Biological Systems Manager, Labcyte, Inc., San Jose, CA
Biological Systems Manager
San Jose, CA
Marsha is a molecular biologist with 10 years of experience in assay product development for basic research (RUO) and molecular diagnostics (CLIA) industries. Passionate about delivering quality products to market on time, with high standards for good science!