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Methods for DNA introduction into mammalian cells
Book chapter   Peer reviewed

Methods for DNA introduction into mammalian cells

Pamela A. Norton and Catherine J. Pachuk
New Comprehensive Biochemistry
2003
DOI: https://doi.org/10.1016/S0167-7306(03)38016-0

Abstract

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The development of methods for the introduction of DNA into cultured cells, especially mammalian cells, has proceeded in parallel with advances in molecular cloning techniques. The process of introducing DNA into vertebrate cells is generally referred to as “transfection,” although some authors have referred to the same process as “transformation,” by analogy with DNA transfer in prokaryotes. In this chapter, the term transfection is used to avoid confusion, as transformation can have a distinct meaning with regard to the growth and the morphologic state of mammalian cells. Ideally, a transfection efficiency of 100% (all target cells acquire and express the introduced DNA) associated with minimal toxicity (all cells survive the procedure) is desired. Although a number of transfection methods have been described, virtually all fall short of these ideals. Increased transfection efficiency often correlates with increased toxicity, necessitating a tradeoff. This chapter reviews the most commonly encountered methods, with consideration of their relative merits both in principle and in practice. The principal barriers to successful transfection are identified and discussed, as further research in this area will likely lead to improved methods in the future. Transfection methods can be grouped simplistically into three categories: physically mediated delivery, chemically mediated delivery and biological vector-mediated delivery of nucleic acid. Although mammalian cells have been transfected with nucleic acids from a variety of sources, including total genomic DNA and RNA, the vast majority of experiments involve DNA sequences that have been subcloned and propagated in Escherichia coli.

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