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Ely, Zackery A; Mathey-Andrews, Nicolas; Naranjo, Santiago; Gould, Samuel I; Mercer, Kim L; Newby, Gregory A; Cabana, Christina M; Rideout, 3rd, William M; Jaramillo, Grissel Cervantes; Khirallah, Jennifer M; Holland, Katie; Randolph, Peyton B; Freed-Pastor, William A; Davis, Jessie R; Kulstad, Zachary; Westcott, Peter M K; Lin, Lin; Anzalone, Andrew V; Horton, Brendan L; Pattada, Nimisha B; Shanahan, Sean-Luc; Ye, Zhongfeng; Spranger, Stefani; Xu, Qiaobing; Sánchez-Rivera, Francisco J; Liu, David R; Jacks, Tyler
Nature biotechnology, 03/2024, Volume: 42, Issue: 3Journal Article
Genetically engineered mouse models only capture a small fraction of the genetic lesions that drive human cancer. Current CRISPR-Cas9 models can expand this fraction but are limited by their reliance on error-prone DNA repair. Here we develop a system for in vivo prime editing by encoding a Cre-inducible prime editor in the mouse germline. This model allows rapid, precise engineering of a wide range of mutations in cell lines and organoids derived from primary tissues, including a clinically relevant Kras mutation associated with drug resistance and Trp53 hotspot mutations commonly observed in pancreatic cancer. With this system, we demonstrate somatic prime editing in vivo using lipid nanoparticles, and we model lung and pancreatic cancer through viral delivery of prime editing guide RNAs or orthotopic transplantation of prime-edited organoids. We believe that this approach will accelerate functional studies of cancer-associated mutations and complex genetic combinations that are challenging to construct with traditional models.
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