title: Modeling Cancer-Associated Mutations of PTEN in Mice.
abstract: Manipulation of mammalian genome and generation of genetically engineered mouse models (GEMMs) has revolutionized the scientific approach to address biological questions. To date, a number of gene-targeting strategies have been devised and are available to investigators for the generation of genetically modified mouse lines. Nevertheless, irrespective of the methodological approach selected, there remain critical molecular steps that need to be performed and put in place in order to obtain controlled and well-characterized new animal models. Here we provide technical details for the (1) handling and maintenance of mouse embryonic stem (ES) cells; (2) analysis of genomic DNA by Southern Blot; and (3) sequencing and PCR analysis of recombined genomic DNA. These experimental steps have been undertaken for the generation of new mouse models harboring cancer-associated PTEN mutations.
authors.affiliation: Monash Biomedicine Discovery Institute and Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC, 3800, Australia. antonella.papa@monash.edu.
authors.forename: Antonella
authors.initials: A
authors.lastname: Papa
authors.affiliation: Cancer Research Institute, Beth Israel Deaconess Cancer Center, Departments of Medicine and Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, 3 Blackfan Circle, Boston, MA, 02215, USA.
authors.forename: Pier Paolo
authors.initials: PP
authors.lastname: Pandolfi
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D055879:Gene Knock-In Techniques / Q000379:methods*; D051379:Mice; D009154:Mutation; D009369:Neoplasms / Q000235:genetics*; D051059:PTEN Phosphohydrolase / Q000235:genetics*
chemical_list: D051059:PTEN Phosphohydrolase; C487757:Pten protein, mouse
keywords: DNA sequencing; Embryonic stem cells; Knock-in mice; PTEN; Point mutations; Southern blot analysis