Nonviral CRISPR/Cas9 mutagenesis for streamlined generation of mouse lung cancer models

• 成果类型: Article
• 署名作者: Lara-Saez, Irene; Mencia, Angeles; Recuero, Enrique; Li, Yinghao; Garcia, Marta; Oteo, Marta; Gallego, Marta I.; Enguita, Ana Belen; de Prado-Verdun, Diana; Sigen, A.; Wang, Wenxin; Garcia-Escuderob, Ramon; Murillas, Rodolfo; Santos, Mirentxu
• 署名单位: University College Dublin; Centro de Investigaciones Energeticas, Medioambientales Tecnologicas; CIBER - Centro de Investigacion Biomedica en Red; CIBERER; Fundacion Jimenez Diaz; Universidad Carlos III de Madrid; Centro de Investigaciones Energeticas, Medioambientales Tecnologicas; Instituto de Salud Carlos III; Hospital Universitario 12 de Octubre; Anhui University of Science & Technology; CIBER - Centro de Investigacion Biomedica en Red; CIBERONC
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-9765
• DOI: 10.1073/pnas.2322917121
• 发表日期: 2024-07-09
• 关键词: cell; genome; delivery; origin; neuroendocrine; inactivation; library; genes; trp53

摘要

Functional analysis in mouse models is necessary to establish the involvement of a set of genetic variations in tumor development. A modeling platform to facilitate and cost- effectively analyze the role of multiple genes in carcinogenesis would be valuable. Here, we present an innovative strategy for lung mutagenesis using CRISPR/Cas9 ribonucleoproteins delivered via cationic polymers. This approach allows the simultaneous inactivation of multiple genes. We validate the effectiveness of this system by targeting a group of tumor suppressor genes, specifically Rb1, Rbl1, Pten, and Trp53, which were chosen for their potential to cause lung tumors, namely small cell lung carcinoma (SCLC). Tumors with histologic and transcriptomic features of human SCLC emerged after intratracheal administration of CRISPR/polymer nanoparticles. These tumors carried loss- of- function mutations in all four tumor suppressor genes at the targeted positions. These findings were reproduced in two different pure genetic backgrounds. We provide a proof of principle for simplified modeling of lung tumorigenesis to facilitate functional testing of potential cancer- related genes.