Modeling Cancer-Associated Mutations of PTEN in Mice

Antonella Papa, Pier Paolo Pandolfi

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

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.
Original languageEnglish
Title of host publicationPTEN
Subtitle of host publicationMethods and Protocols
EditorsLeonardo Salmena, Vuk Stambolic
Place of PublicationNew York NY USA
PublisherHumana Press
Pages289-306
Number of pages18
Edition1st
ISBN (Electronic)978-1-4939-3299-3
ISBN (Print)978-1-4939-3297-9
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology: Springer Protocols
PublisherSpringer
Volume1388
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

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