Modelling intestinal carcinogenesis using in vitro organoid cultures

Thierry Jardé, Genevieve Kerr, Reyhan Akhtar, Helen E. Abud

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

Abstract

Mouse models of intestinal carcinogenesis are very powerful for studying the impact of specific mutations on tumour initiation and progression. Mutations can be studied both singularly and in combination using conditional alleles that can be induced in a temporal manner. The steps in intestinal carcinogenesis are complex and can be challenging to image in live animals at a cellular level. The ability to culture intestinal epithelial tissue in three-dimensional organoids in vitro provides an accessible system that can be genetically manipulated and easily visualised to assess specific biological impacts in living tissue. Here, we describe methodology for conditional mutation of genes in organoids from genetically modified mice via induction of Cre recombinase induced by tamoxifen or by transient exposure to TAT-Cre protein. This methodology provides a rapid platform for assessing the cellular changes induced by specific mutations in intestinal tissue.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Pages41-52
Number of pages12
Volume1725
DOIs
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1725
ISSN (Print)1064-3745

Keywords

  • Conditional mutation
  • Cre recombinase
  • Intestinal organoids

Cite this

Jardé, T., Kerr, G., Akhtar, R., & Abud, H. E. (2018). Modelling intestinal carcinogenesis using in vitro organoid cultures. In Methods in Molecular Biology (Vol. 1725, pp. 41-52). (Methods in Molecular Biology; Vol. 1725). Humana Press. https://doi.org/10.1007/978-1-4939-7568-6_4
Jardé, Thierry ; Kerr, Genevieve ; Akhtar, Reyhan ; Abud, Helen E. / Modelling intestinal carcinogenesis using in vitro organoid cultures. Methods in Molecular Biology. Vol. 1725 Humana Press, 2018. pp. 41-52 (Methods in Molecular Biology).
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Jardé, T, Kerr, G, Akhtar, R & Abud, HE 2018, Modelling intestinal carcinogenesis using in vitro organoid cultures. in Methods in Molecular Biology. vol. 1725, Methods in Molecular Biology, vol. 1725, Humana Press, pp. 41-52. https://doi.org/10.1007/978-1-4939-7568-6_4

Modelling intestinal carcinogenesis using in vitro organoid cultures. / Jardé, Thierry; Kerr, Genevieve; Akhtar, Reyhan; Abud, Helen E.

Methods in Molecular Biology. Vol. 1725 Humana Press, 2018. p. 41-52 (Methods in Molecular Biology; Vol. 1725).

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

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AB - Mouse models of intestinal carcinogenesis are very powerful for studying the impact of specific mutations on tumour initiation and progression. Mutations can be studied both singularly and in combination using conditional alleles that can be induced in a temporal manner. The steps in intestinal carcinogenesis are complex and can be challenging to image in live animals at a cellular level. The ability to culture intestinal epithelial tissue in three-dimensional organoids in vitro provides an accessible system that can be genetically manipulated and easily visualised to assess specific biological impacts in living tissue. Here, we describe methodology for conditional mutation of genes in organoids from genetically modified mice via induction of Cre recombinase induced by tamoxifen or by transient exposure to TAT-Cre protein. This methodology provides a rapid platform for assessing the cellular changes induced by specific mutations in intestinal tissue.

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Jardé T, Kerr G, Akhtar R, Abud HE. Modelling intestinal carcinogenesis using in vitro organoid cultures. In Methods in Molecular Biology. Vol. 1725. Humana Press. 2018. p. 41-52. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-7568-6_4