In vivo genome editing and organoid transplantation models of colorectal cancer and metastasis.

Published

Journal Article

In vivo interrogation of the function of genes implicated in tumorigenesis is limited by the need to generate and cross germline mutant mice. Here we describe approaches to model colorectal cancer (CRC) and metastasis, which rely on in situ gene editing and orthotopic organoid transplantation in mice without cancer-predisposing mutations. Autochthonous tumor formation is induced by CRISPR-Cas9-based editing of the Apc and Trp53 tumor suppressor genes in colon epithelial cells and by orthotopic transplantation of Apc-edited colon organoids. ApcΔ/Δ;KrasG12D/+;Trp53Δ/Δ (AKP) mouse colon organoids and human CRC organoids engraft in the distal colon and metastasize to the liver. Finally, we apply the orthotopic transplantation model to characterize the clonal dynamics of Lgr5+ stem cells and demonstrate sequential activation of an oncogene in established colon adenomas. These experimental systems enable rapid in vivo characterization of cancer-associated genes and reproduce the entire spectrum of tumor progression and metastasis.

Full Text

Duke Authors

Cited Authors

  • Roper, J; Tammela, T; Cetinbas, NM; Akkad, A; Roghanian, A; Rickelt, S; Almeqdadi, M; Wu, K; Oberli, MA; Sánchez-Rivera, FJ; Park, YK; Liang, X; Eng, G; Taylor, MS; Azimi, R; Kedrin, D; Neupane, R; Beyaz, S; Sicinska, ET; Suarez, Y; Yoo, J; Chen, L; Zukerberg, L; Katajisto, P; Deshpande, V; Bass, AJ; Tsichlis, PN; Lees, J; Langer, R; Hynes, RO; Chen, J; Bhutkar, A; Jacks, T; Yilmaz, ÖH

Published Date

  • June 2017

Published In

Volume / Issue

  • 35 / 6

Start / End Page

  • 569 - 576

PubMed ID

  • 28459449

Pubmed Central ID

  • 28459449

Electronic International Standard Serial Number (EISSN)

  • 1546-1696

Digital Object Identifier (DOI)

  • 10.1038/nbt.3836

Language

  • eng

Conference Location

  • United States