E-Book Overview
This superb collection of well-tested protocols for the electroporation of proteins and DNA into insect, fish, and mammalian cells also includes proven animal cell electrofusion techniques for studies of somatic cell genetics and development, and for generating monoclonal antibodies. It is distinguished by its coverage of important model cell types from many organisms and tissue types and its detailed instructions for the growth and preparation of specific cells to achieve optimum transfection. Its extensive reference lists, citations of alternative transfer methods, advice on pitfalls to avoid, and descriptions of expected results ensure readily reproducible results even for beginners.
E-Book Content
CHAFFER 1 Electroporation Theory Concepts and Mechanisms James C. Weaver 1. Introduction Application of strong electric field pulses to cells and tissue is known to cause some type of structural rearrangement of the cell membrane. Significant progress has been made by adopting the hypothesis that some of these rearrangements consist of temporary aqueous pathways (“pores”), with the electric field playing the dual role of causing pore formation and providing a local driving force for ionic and molecular transport through the pores. Introduction of DNA into cells in vitro is now the most common application. With imagination, however, many other uses seem likely. For example, in vitro electroporation has been used to introduce into cells enzymes, antibodies, and other biochemical reagents for intracellular assays; to load larger cells preferentially with molecules in the presence of many smaller cells; to introduce particles into cells, including viruses; to kill cells purposefully under otherwise mild conditions; and to insert membranemacromolecules into the cell membrane itself. Only recently has the exploration of in vivo electroporation for