| Literature DB >> 25924068 |
Jarno Drost1, Richard H van Jaarsveld2, Bas Ponsioen2, Cheryl Zimberlin3, Ruben van Boxtel1, Arjan Buijs4, Norman Sachs1, René M Overmeer2, G Johan Offerhaus5, Harry Begthel1, Jeroen Korving1, Marc van de Wetering6, Gerald Schwank1, Meike Logtenberg1, Edwin Cuppen1, Hugo J Snippert2, Jan Paul Medema3, Geert J P L Kops2, Hans Clevers1.
Abstract
Crypt stem cells represent the cells of origin for intestinal neoplasia. Both mouse and human intestinal stem cells can be cultured in medium containing the stem-cell-niche factors WNT, R-spondin, epidermal growth factor (EGF) and noggin over long time periods as epithelial organoids that remain genetically and phenotypically stable. Here we utilize CRISPR/Cas9 technology for targeted gene modification of four of the most commonly mutated colorectal cancer genes (APC, P53 (also known as TP53), KRAS and SMAD4) in cultured human intestinal stem cells. Mutant organoids can be selected by removing individual growth factors from the culture medium. Quadruple mutants grow independently of all stem-cell-niche factors and tolerate the presence of the P53 stabilizer nutlin-3. Upon xenotransplantation into mice, quadruple mutants grow as tumours with features of invasive carcinoma. Finally, combined loss of APC and P53 is sufficient for the appearance of extensive aneuploidy, a hallmark of tumour progression.Entities:
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Year: 2015 PMID： 25924068 DOI： 10.1038/nature14415
Source DB: PubMed Journal: Nature ISSN： 0028-0836 Impact factor: 49.962