title: Tamoxifen-inducible Cre-mediated calreticulin excision to study mouse embryonic stem cell differentiation.
abstract: Embryonic stem cells are useful to study the functional aspects of lineage commitment. In this study, we report that using the Cre/loxP system provides a useful tool for studying multifunctional proteins that are involved in stem cell differentiation, such as calreticulin. Calreticulin is a chaperone and a major calcium buffer of the endoplasmic reticulum and it functions during both adipogenesis and cardiomyogenesis. We used both a tamoxifen-inducible and cardiomyocyte-specific alpha-myosin heavy chain promoter-driven Cre/loxP system to study cardiomyogenesis, and a tamoxifen-inducible ubiquitously expressed cytomegalovirus promoter-driven Cre/loxP system to study adipogenesis. Both Cre/loxP systems mimicked the results previously observed using the calreticulin-null stem cell systems. Our results indicate that the tamoxifen-inducible Cre/loxP system is an effective and reliable tool to use for gene ablation in studies on functional aspects of stem cell biology.
authors.affiliation: Laboratory Medicine and Pathobiology/Institute of Medical Sciences, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Eva
authors.initials: E
authors.lastname: Szabo
authors.forename: Jonathan
authors.initials: J
authors.lastname: Soboloff
authors.forename: Ewa
authors.initials: E
authors.lastname: Dziak
authors.forename: Michal
authors.initials: M
authors.lastname: Opas
mesh_terms: D000818:Animals; D037282:Calreticulin / Q000235:genetics / Q000378:metabolism*; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000502:physiology; D011401:Promoter Regions, Genetic; D013629:Tamoxifen / Q000378:metabolism*
chemical_list: D037282:Calreticulin; D013629:Tamoxifen