Stem cells engendered with cassette exchange system- approach to study gene gain of function

Bosnakovski, Darko (2012) Stem cells engendered with cassette exchange system- approach to study gene gain of function. In: 2nd Joint Meeting; From nano to macro biomaterials (design, processing, characterization, modeling) and applications to stem cells regenerative orthopedic and dental medicine (NAMABIO).

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Abstract

Stem cells engendered with cassette exchange system- approach to study gene gain of function Darko Bosnakovski1,2* , Michelina Iacovino2, Holger Fey2 , Ana Mitanoska1,2 and Michael Kyba2 1University Goce Delčev - Štip, Faculty of Medical Sciences, Krste Misirkov b.b., 2000 Štip, R. Macedonia 2Lillehei Heart Institute, Department of Pediatrics, University of Minnesota, Minneapolis, MN, 55014 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 in and reprogramming of cell fate. REFERENCES: Bosnakovski D, Xu Z, Gang EJ, Galindo CL, Liu M, Simsek T, Garner HR, Agha-Mohammadi S, Tassin A, Coppee F et al: An isogenetic myoblast expression screen identifies DUX4-mediated FSHD-associated molecular pathologies. EMBO J 2008, 27(20):2766-2779. Kyba M, Perlingeiro RC, Daley GQ: HoxB4 confers definitive lymphoid-myeloid engraftment potential on embryonic stem cell and yolk sac hematopoietic progenitors. Cell 2002, 109(1):29-37 .

Item Type: Conference or Workshop Item (Poster)
Subjects: Natural sciences > Biological sciences
Medical and Health Sciences > Health biotechnology
Divisions: Faculty of Medical Science
Depositing User: Darko Bosnakovski
Date Deposited: 05 Feb 2013 15:36
Last Modified: 05 Feb 2013 15:36
URI: http://eprints.ugd.edu.mk/id/eprint/5046

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