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Rapid interrogation of cancer cell of origin through CRISPR editing.

Proceedings of the National Academy of Sciences of the United States of America (2021-08-07)
Weiran Feng, Zhen Cao, Pei Xin Lim, Huiyong Zhao, Hanzhi Luo, Ninghui Mao, Young Sun Lee, Aura Agudelo Rivera, Danielle Choi, Chao Wu, Teng Han, Rodrigo Romero, Elisa de Stanchina, Brett S Carver, Qiao Wang, Maria Jasin, Charles L Sawyers
ABSTRACT

The increasing complexity of different cell types revealed by single-cell analysis of tissues presents challenges in efficiently elucidating their functions. Here we show, using prostate as a model tissue, that primary organoids and freshly isolated epithelial cells can be CRISPR edited ex vivo using Cas9-sgRNA (guide RNA) ribotnucleoprotein complex technology, then orthotopically transferred in vivo into immunocompetent or immunodeficient mice to generate cancer models with phenotypes resembling those seen in traditional genetically engineered mouse models. Large intrachromosomal (∼2 Mb) or multigenic deletions can be engineered efficiently without the need for selection, including in isolated subpopulations to address cell-of-origin questions.

MATERIALS
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Product Description

Sigma-Aldrich
Anti-APC Antibody, clone FE9, clone FE9, from mouse