High‐throughput ‘On Chip’ Protein and Nucleic Acid Transfection

Email a friend
Contact the editor about this article
How to cite

High‐throughput ‘On Chip’ Protein and Nucleic Acid Transfection

Frank Herrmann, Centre for Genomic Regulation (CRG), Barcelona, Spain
Mark Isalan, Centre for Genomic Regulation (CRG), Barcelona, Spain

Published online: July 2008

DOI: 10.1002/9780470015902.a0020899

Full Article on Wiley Online Library

Abstract
Images
References

Transfection describes the nonviral introduction of exogenous molecules into eukaryotic cells. Several transfection methods have been developed to transfer nucleic acids, proteins and other macromolecules across the plasma membrane efficiently. To facilitate large-scale genomic and proteomic studies there is an increasing need for automated high-throughput transfection platforms.

Keywords: transfection; protein transfection; reverse transfection; cell microarray

References
	other Fang Y, Lai F, Picard LAG and Webb BL (2006) Method and device for protein delivery into cells. United States Patent 7105347.
	Felgner PL, Tsai YJ, Sukhu L et al. (1995) Improved cationic lipid formulations for in vivo gene therapy. Annals of the New York Academy of Sciences 772: 126–139.
	Fox MB, Esveld DC, Valero A et al. (2006) Electroporation of cells in microfluidic devices: a review. Analytical and Bioanalytical Chemistry 385: 474–485.
	Frankel AD and Pabo CO (1988) Cellular uptake of the tat protein from human immunodeficiency virus. Cell 55: 1189–1193.
	Green M and Loewenstein PM (1988) Autonomous functional domains of chemically synthesized human immunodeficiency virus tat trans-activator protein. Cell 55: 1179–1188.
	Isalan M, Santori MI, Gonzalez C and Serrano L (2005) Localized transfection on arrays of magnetic beads coated with PCR products. Nature Methods 2: 113–118.
	Jain T and Muthuswamy J (2007) Bio-chip for spatially controlled transfection of nucleic acid payloads into cells in a culture. Lab on a Chip 7: 1004–1011.
	Lambert H, Pankov R, Gauthier J and Hancock R (1990) Electroporation-mediated uptake of proteins into mammalian cells. Biochemistry and Cell Biology 68: 729–734.
	Lin Y-C, Jen C-M, Huang M-Y, Wu C-Y and Lin X-Z (2001) Electroporation microchips for continuous gene transfection. Sensors and Actuators B: Chemical 79: 137–143.
	Matsuoka H, Komazaki T, Mukai Y et al. (2005) High throughput easy microinjection with a single-cell manipulation supporting robot. Journal of Biotechnology 116: 185–194.
	Morris MC, Depollier J, Mery J, Heitz F and Divita G (2001) A peptide carrier for the delivery of biologically active proteins into mammalian cells. Nature Biotechnology 19: 1173–1176.
	Neumann B, Held M, Liebel U et al. (2006) High-throughput RNAi screening by time-lapse imaging of live human cells. Nature Methods 3: 385–390.
	Okazaki A, Jo J and Tabata Y (2007) A reverse transfection technology to genetically engineer adult stem cells. Tissue Engineering 13: 245–251.
	Palmer EL, Miller AD and Freeman TC (2006) Identification and characterisation of human apoptosis inducing proteins using cell-based transfection microarrays and expression analysis. BMC Genomics 7: 145.
	Santori MI, Gonzalez C, Serrano L and Isalan M (2006) Localized transfection with magnetic beads coated with PCR products and other nucleic acids. Nature Protocols 1: 526–531.
	Scherer F, Anton M, Schillinger U et al. (2002) Magnetofection: enhancing and targeting gene delivery by magnetic force in vitro and in vivo. Gene Therapy 9: 102–109.
	Wheeler DB, Bailey SN, Guertin DA et al. (2004) RNAi living-cell microarrays for loss-of-function screens in Drosophila melanogaster cells. Nature Methods 1: 127–132.
	Ziauddin J and Sabatini DM (2001) Microarrays of cells expressing defined cDNAs. Nature 411: 107–110.
Further Reading
	book Sambrook J, Russell DW (2001) "Introducing cloned genes in cultured mammalian cells". In: Argentine J and Irwin N (eds) Molecular Cloning: A Laboratory Manual, chap. 16. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.
	Starkuviene V, Pepperkok R and Erfle H (2007) Transfected cell microarrays: an efficient tool for high-throughput functional analysis. Expert Review of Proteomics 4: 479–489.
	Valero A, Post JN, van Nieuwkasteele JW et al. (2008) Gene transfer and protein dynamics in stem cells using single cell electroporation in a microfluidic device. Lab on a Chip 8: 62–67.

Article Title:	High‐throughput ‘On Chip’ Protein and Nucleic Acid Transfection
* Name:
* E-mail:
* Note:

	Figure 1. Overview of methods of delivery of nucleic acids and proteins across cell membranes.
	Figure 2. The bead transfection method (Isalan et al., 2005). Streptavidin-coated Dynabeads (3 m diameter) bind to the biotinylated molecules that are to be transfected (e.g. PCR DNA, siRNA or protein). Coated beads are incubated with transfection reagent and exposed to the cells. In the example to the right, a PCR product encoding the EGFP gene under a CMV promoter is made with a biotinylated primer; a single bead is shown transfecting a single cell within a lawn of MDCK cells.
	Figure 3. The principle of reverse transfection microarrays (Ziauddin and Sabatini, 2001). The transfected macromolecule is spotted onto a glass slide and is subsequently coated with transfection reagent and then cells. The spots result in locally transfected fields of cells which can be analysed by microscopy.

High‐throughput ‘On Chip’ Protein and Nucleic Acid Transfection

Related Sub-Topics: