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

Cite this

Papa, A., & Pandolfi, P. P. (2016). Modeling Cancer-Associated Mutations of PTEN in Mice. In L. Salmena, & V. Stambolic (Eds.), PTEN: Methods and Protocols (1st ed., pp. 289-306). (Methods in Molecular Biology: Springer Protocols; Vol. 1388). New York NY USA: Humana Press. https://doi.org/10.1007/978-1-4939-3299-3_16
Papa, Antonella ; Pandolfi, Pier Paolo. / Modeling Cancer-Associated Mutations of PTEN in Mice. PTEN: Methods and Protocols. editor / Leonardo Salmena ; Vuk Stambolic. 1st. ed. New York NY USA : Humana Press, 2016. pp. 289-306 (Methods in Molecular Biology: Springer Protocols).
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Papa, A & Pandolfi, PP 2016, Modeling Cancer-Associated Mutations of PTEN in Mice. in L Salmena & V Stambolic (eds), PTEN: Methods and Protocols. 1st edn, Methods in Molecular Biology: Springer Protocols, vol. 1388, Humana Press, New York NY USA, pp. 289-306. https://doi.org/10.1007/978-1-4939-3299-3_16

Modeling Cancer-Associated Mutations of PTEN in Mice. / Papa, Antonella; Pandolfi, Pier Paolo.

PTEN: Methods and Protocols. ed. / Leonardo Salmena; Vuk Stambolic. 1st. ed. New York NY USA : Humana Press, 2016. p. 289-306 (Methods in Molecular Biology: Springer Protocols; Vol. 1388).

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

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AB - 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.

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Papa A, Pandolfi PP. Modeling Cancer-Associated Mutations of PTEN in Mice. In Salmena L, Stambolic V, editors, PTEN: Methods and Protocols. 1st ed. New York NY USA: Humana Press. 2016. p. 289-306. (Methods in Molecular Biology: Springer Protocols). https://doi.org/10.1007/978-1-4939-3299-3_16