title: Targeting iCre expression to murine progesterone receptor cell-lineages using bacterial artificial chromosome transgenesis.
abstract: Gene-targeting in embryonic stem cells has been the dominant genetic approach when engineering mouse models to query the physiologic importance of the progesterone receptor (PR). Although these models have been instrumental in disclosing the in vivo significance of the progesterone signaling pathway, generation of such mice exacts considerable expenditure of time, effort, and expense. Considering the growing list of new PR mouse models that are urgently required to address the next questions in progestin biology, bacterial artificial chromosome (BAC) recombineering in conjunction with transgenesis was evaluated as an alternative method to accelerate the creation of these models in the future. Using this approach, we describe the generation of three PR-BAC(iCre) transgenic lines in which improved Cre recombinase (iCre) was targeted in-frame, downstream, and under the control of the PR promoter contained within a BAC transgene. Crossing with the ROSA26R revealed that the PR-BAC(iCre) transgenic expresses active iCre only in cell-lineages that express the PR. The specificity of the PR-BAC(iCre) transgene not only underscores the importance of BAC-mediated transgenesis as a quick, easy, and affordable method by which to engineer the next generation of PR mouse models, but also provides a unique opportunity to investigate transcriptional control of PR expression as well as PR structure-function relationships in vivo.
authors.affiliation: Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: Atish
authors.initials: A
authors.lastname: Mukherjee
authors.forename: Selma M
authors.initials: SM
authors.lastname: Soyal
authors.forename: David A
authors.initials: DA
authors.lastname: Wheeler
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Fernandez-Valdivia
authors.forename: Jonathan
authors.initials: J
authors.lastname: Nguyen
authors.forename: Francesco J
authors.initials: FJ
authors.lastname: DeMayo
authors.forename: John P
authors.initials: JP
authors.lastname: Lydon
mesh_terms: D000818:Animals; D019070:Cell Lineage / Q000235:genetics*; D022202:Chromosomes, Artificial, Bacterial; D019295:Computational Biology; D017931:DNA Primers; D040461:Gene Components; D018014:Gene Transfer Techniques; D019426:Integrases / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D023421:Models, Animal; D011401:Promoter Regions, Genetic / Q000235:genetics; D011980:Receptors, Progesterone / Q000235:genetics* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D011980:Receptors, Progesterone; C045073:Cre recombinase; D019426:Integrases