Authors
Jarno Drost, Richard H Van Jaarsveld, Bas Ponsioen, Cheryl Zimberlin, Ruben Van Boxtel, Arjan Buijs, Norman Sachs, René M Overmeer, G Johan Offerhaus, Harry Begthel, Jeroen Korving, Marc Van De Wetering, Gerald Schwank, Meike Logtenberg, Edwin Cuppen, Hugo J Snippert, Jan Paul Medema, Geert JPL Kops, Hans Clevers
Publication date
2015/5/7
Journal
Nature
Volume
521
Issue
7550
Pages
43-47
Publisher
Nature Publishing Group UK
Description
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 …
Scholar articles
J Drost, RH Van Jaarsveld, B Ponsioen, C Zimberlin… - Nature, 2015