High-throughput ‘On Chip’ Protein and Nucleic Acid Transfection
Published Online: 15 APR 2014
Copyright © 2001 John Wiley & Sons, Ltd. All rights reserved.
How to Cite
Herrmann, F. and Isalan, M. 2014. High-throughput ‘On Chip’ Protein and Nucleic Acid Transfection. eLS. .
- Published Online: 15 APR 2014
Transfection describes the nonviral introduction of exogenous molecules into eukaryotic cells. Numerous transfection methods have been developed to transfer nucleic acids, proteins and other macromolecules across the plasma membrane efficiently. These include physical methods, such as electroporation, magnetofection and microinjection, as well as a chemical or biological carrier-mediated methods. Chemical transfection reagents such as cationic lipids or polymers are widely used, either alone or in combination with scaffolds. Biological methods include delivery with cell-penetrating protein domain fusions such as trans-activator of transcription protein from human immunodeficiency virus, VP22 or Antennapedia peptides. Certain proteins such as zinc-finger nucleases, which are used for targeted genome modification, even appear to have intrinsic cell-penetrating properties. To facilitate large-scale genomic and proteomic studies there is an increasing need for automated high-throughput transfection platforms. Several such platforms have been developed, including multiwell plates, transfection microarrays and microfluidic chip formats. Overall, the different advantages and applications of these diverse methods to deliver cargo into cells are discussed.
Cell membranes are a barrier to the delivery of proteins and nucleic acids into cells.
Mechanical, chemical and biological methods of transfection have been developed to cross the cell membrane barrier.
Positively charged molecules, such as cationic lipids, polymers or peptides, can help to penetrate the negatively-charged surface of cell membranes.
Various automatable high-throughput technologies have been developed, including transfection microarrays.
Reverse transfection is a scalable, potentially high-throughput process where cells are added to transfectable molecules, preimmobilised on a scaffold.
- protein transfection;
- reverse transfection;
- cell microarray;
- gene delivery