Rapid interrogation of cancer cell of origin through CRISPR editing
Abstract
Modeling cancer formation requires introduction of relevant oncogenic perturbations into normal cells in a tissue/cell-type–specific manner. Genetically engineered mouse models are powerful but require significant time and cost to generate and maintain. The ability to edit primary epithelial cells ex vivo followed by orthotopic transplantation provides an alternative strategy for cancer modeling but requires efficient gene editing, typically in a multiplex fashion. Here we successfully engineer multigenic perturbations or chromosomal rearrangements in primary prostate organoids through single-step Cas9–sgRNA ribonucleoprotein electroporation. This approach can also address cell-of-origin questions by directly editing and transplanting freshly isolated subpopulations without the intermediate step of organoid culture, providing a rapid complement to traditional lineage tracing approaches.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Aug 05, 2021
- Source ID
- 10.1073/pnas.2110344118
Entities
People
- Aura Agudelo Rivera
- Brett S. Carver
- Chao Wu
- Charles Sawyers
- Danielle Choi
- Elisa De Stanchina
- Hanzhi Luo
- Huiyong Zhao
- Maria Jasin
- Ninghui Mao
- Pei Xin Lim
- Qiao Wang
- Rodrigo Romero
- Teng Han
- Weiran Feng
- Young Sun Lee
- Zhen Cao
Organizations
- Fudan University
- Howard Hughes Medical Institute
- Memorial Sloan Kettering Cancer Center
- National Cancer Institute
- United States Department of Defense
- Weill Cornell Medicine