SpringerProtocols: Abstract: siRNA and DNA Transfer to Cultured Cells

3. siRNA and DNA Transfer to Cultured Cells

By: Bagavathi Gopalakrishnan¹, Jon Wolff²

Abstract

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Transfection is a powerful non-viral technology used to deliver foreign nucleic acids into eukaryotic cells, and is the method of choice for a variety of applications including studying the functional role of particular genes and the proteins they code for. By over-expressing genes to produce protein of interest and also by knocking down specific genes, researchers are able to accurately define the role of genes and the protein they encode in various cellular processes. Therefore, this powerful technology is a very vital component of the array of scientific research tools. However, the exact mechanism of action of transfection and also the numerous factors that influence the success of DNA or RNA delivery processes are not clearly understood. Hence, this chapter attempts to explain some of the popular cationic lipid/polymer-based transfection reagents for in vitro DNA/small inhibitory RNA (siRNA) delivery, mainly focusing on the protocols and critical factors to keep in mind to ensure successful delivery of nucleic acids into eukaryotic cells using these methods.

Affiliation(s): (1) Mirus Bio Corporation, 505 S. Rosa Road, Madison, WI 53719, USA
(2) Departments of Pediatrics and Medical Genetics, Waisman Center, University of Wisconsin-Madison, Madison, WI 53705, USA

Book Title: Macromolecular Drug Delivery: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 480 | Year: 2009 | Page Range: 1-22 | DOI: 10.1007/978-1-59745-429-2_3

Subject: Pharmacology/Toxicology

Key Words: Transfection - Non-viral DNA delivery - RNA delivery - siRNA delivery - Nucleic acid transfer - Knockdown - Reporter assay

References

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