Receptor-mediated transport of DNA into eukaryotic cells

Cotten, M. , Wagner, E. and Birnstiel, M. L. (1993) Receptor-mediated transport of DNA into eukaryotic cells. Methods in Enzymology, 217, pp. 618-644. (doi:10.1016/0076-6879(93)17092-j) (PMID:8386298)

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Abstract

This chapter describes the methods for preparing and using transferrin-polycation and other ligand-polycation conjugates for receptor-mediated DNA transfer. This method, termed “transferrinfection,” is particularly effective with cell lines derived from the erythroid lineage most likely because of the high level of cycling transferrin receptor on these cells. In other established cell lines—such as HeLa, CHO, Cos, and HepG2—this method works with efficiencies comparable to other transfection techniques. In the cells where transferrinfection functions well, the method has the following advantages over other transfection methods: (1) the method is simple to use, once the ligand-polycation conjugates are obtained, (2) the method can be used with many DNA molecules, and (3) the method is particularly gentle, involving a transferrin-polycation conjugate that the cell binds with nearly the same avidity as unmodified transferrin. Fluorescence-activated cell sorting (FACS) analyses of transfected cell populations demonstrate >95% viability at 24 hr, 48 hr, and 7 days posttransfection. This may partly account for the high efficiencies obtained.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Cotten, Professor Matthew
Authors: Cotten, M., Wagner, E., and Birnstiel, M. L.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Methods in Enzymology
Publisher:Academic Press
ISSN:0076-6879
ISSN (Online):1557-7988

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