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Publication : Inducible cassette exchange: a rapid and efficient system enabling conditional gene expression in embryonic stem and primary cells.

First Author  Iacovino M Year  2011
Journal  Stem Cells Volume  29
Issue  10 Pages  1580-8
PubMed ID  22039605 Mgi Jnum  J:212531
Mgi Id  MGI:5581705 Doi  10.1002/stem.715
Citation  Iacovino M, et al. (2011) Inducible cassette exchange: a rapid and efficient system enabling conditional gene expression in embryonic stem and primary cells. Stem Cells 29(10):1580-8
abstractText  Genetic modification is critically enabling for studies addressing specification and maintenance of cell fate; however, methods for engineering modifications are inefficient. We demonstrate a rapid and efficient recombination system in which an inducible, floxed cre allele replaces itself with an incoming transgene. We target this inducible cassette exchange (ICE) allele to the (HPRT) locus and demonstrate recombination in murine embryonic stem cells (ESCs) and primary cells from derivative ICE mice. Using lentivectors, we demonstrate recombination at a randomly integrated ICE locus in human ESCs. To illustrate the utility of this system, we insert the myogenic regulator, Myf5, into the ICE locus in each platform. This enables efficient directed differentiation of mouse and human ESCs into skeletal muscle and conditional myogenic transdetermination of primary cells cultured in vitro. This versatile tool is thus well suited to gain-of-function studies probing gene function in the specification and reprogramming of cell fate.
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