Transgenic cardiomyocytes with controlled proliferation and differentiation
AssigneeBoard of Regents, The University of Texas System
InventorsEric Olson, Rhonda Bassel-Duby, David W. Markham, Igor I. Rybkin, R. Sanders Williams
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Abstract
The present invention provides methods for creating conditionally-immortal cell lines. These transgenic cell lines can be grown indefinitely in culture while maintaining a relatively undifferentiated stated. Upon appropriate switch signal, the cells cease replicating and differentiate much like adult cells. The switch is facilitated by the inactivation of a transforming gene, such as large T antigen. A convenient methodology for such inactivation is Cre-Lox mediated excision of the gene. Cardiac cells are provided as an example of useful a transgenic cell line.
Claims22
A transgenic mouse, cells of which comprise an expression cassette comprising a tissue selective promoter operably linked to a nucleic acid segment encoding SV40 large T antigen, wherein said nucleic acid segment is flanked 5′ and 3′ by site specific excision sequences.
A method for obtaining a transgenic murine progenitor cell line comprising:
- (a) transforming one or more murine embryonic cells with an expression cassette comprising a tissue selective promoter operably linked to a nucleic acid segment encoding SV40 large T antigen, wherein said nucleic acid segment is flanked 5′ and 3′ by site specific excision sequences;
- (b) inserting said one or more murine embryonic cells into a surrogate mouse mother;
- (c) obtaining one or more pups from said surrogate mouse mother;
- (d) identifying one or more pups that express SV40 large T antigen in a tissue selective manner;
- and
- (e) obtaining cells from said one or more pups that express SV40 large T antigen.
A transgenic murine progenitor cell line comprising an expression cassette comprising a tissue selective promoter operably linked to a nucleic acid segment encoding SV40 large T antigen, wherein said nucleic acid segment is flanked 5′ and 3′ by site specific excision sequences.
Dependencies are inferred from the claim text.