Gene Transfer to the Rodent Embryo by Retroviral Vectors
Over the last 20 years investigators studying vertebrate development have capitalized on the use of retroviral-mediated gene transfer to determine the lineage relationships of diverse cell types, particularly in many regions of the mammalian central nervous system (for review, see ref. 1). Whereas an intraperitoneal or intrauterine injection of cell proliferation markers such as tritiated thymidine or bromodeoxyuridine suffices to examine the birthdates of cells, many fundamental questions dealing with the formation of a structure in the mammalian embryo often necessitate performing intrauterine surgery to introduce genes by retroviral or other viral vectors. As retroviral vectors can be used not only to study lineage (2), but also to introduce genes to perturb development (3, 4, 5), the methods for delivering retroviruses or other viruses into the developing mammalian embryo will be in increasing demand. This chapter describes a set of procedures to generate and...
KeywordsChloral Hydrate Xiphoid Process Packaging Cell Line Acetate Sheet Biological Safety Cabinet
We thank Dr. John Parnavelas for advice pertaining to electron microscopy. The authors—research has been supported in part by grants from the National Institutes of Health to M. B. L (#NS28380) and to G. K. P (#AR43410) and by a Biomedical Scholars Award from the Pew Charitable Trusts and a Basic Research Grant from the March of Dimes Birth Defects Foundation to M. B. L.
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