Open AccessRapid interrogation of cancer cell of origin through CRISPR editing
Author(s) -
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
Publication year - 2021
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.2110344118
Subject(s) - crispr , biology , ex vivo , computational biology , genome editing , guide rna , subgenomic mrna , cas9 , cell , phenotype , in vivo , microbiology and biotechnology , cancer cell , cancer , genetics , gene
Significance 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.