Implementation of CRISPR/Cas9 Genome Editing to Generate Murine Lung Cancer Models That Depict the Mutational Landscape of Human Disease

Please always quote using this URN: urn:nbn:de:bvb:20-opus-230949
  • Lung cancer is the most common cancer worldwide and the leading cause of cancer-related deaths in both men and women. Despite the development of novel therapeutic interventions, the 5-year survival rate for non-small cell lung cancer (NSCLC) patients remains low, demonstrating the necessity for novel treatments. One strategy to improve translational research is the development of surrogate models reflecting somatic mutations identified in lung cancer patients as these impact treatment responses. With the advent of CRISPR-mediated genomeLung cancer is the most common cancer worldwide and the leading cause of cancer-related deaths in both men and women. Despite the development of novel therapeutic interventions, the 5-year survival rate for non-small cell lung cancer (NSCLC) patients remains low, demonstrating the necessity for novel treatments. One strategy to improve translational research is the development of surrogate models reflecting somatic mutations identified in lung cancer patients as these impact treatment responses. With the advent of CRISPR-mediated genome editing, gene deletion as well as site-directed integration of point mutations enabled us to model human malignancies in more detail than ever before. Here, we report that by using CRISPR/Cas9-mediated targeting of Trp53 and KRas, we recapitulated the classic murine NSCLC model Trp53fl/fl:lsl-KRasG12D/wt. Developing tumors were indistinguishable from Trp53fl/fl:lsl-KRasG12D/wt-derived tumors with regard to morphology, marker expression, and transcriptional profiles. We demonstrate the applicability of CRISPR for tumor modeling in vivo and ameliorating the need to use conventional genetically engineered mouse models. Furthermore, tumor onset was not only achieved in constitutive Cas9 expression but also in wild-type animals via infection of lung epithelial cells with two discrete AAVs encoding different parts of the CRISPR machinery. While conventional mouse models require extensive husbandry to integrate new genetic features allowing for gene targeting, basic molecular methods suffice to inflict the desired genetic alterations in vivo. Utilizing the CRISPR toolbox, in vivo cancer research and modeling is rapidly evolving and enables researchers to swiftly develop new, clinically relevant surrogate models for translational research.show moreshow less

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Author: Oliver Hartmann, Michaela Reissland, Carina R. Maier, Thomas Fischer, Cristian Prieto-Garcia, Apoorva Baluapuri, Jessica Schwarz, Werner Schmitz, Martin Garrido-Rodriguez, Nikolett Pahor, Clare C. Davies, Florian Bassermann, Amir Orian, Elmar Wolf, Almut Schulze, Marco A. Calzado, Mathias T. Rosenfeldt, Markus E. Diefenbacher
URN:urn:nbn:de:bvb:20-opus-230949
Document Type:Journal article
Faculties:Medizinische Fakultät / Klinik und Poliklinik für Strahlentherapie
Medizinische Fakultät / Pathologisches Institut
Medizinische Fakultät / Theodor-Boveri-Institut für Biowissenschaften
Fakultät für Biologie / Theodor-Boveri-Institut für Biowissenschaften
Language:English
Parent Title (English):Frontiers in Cell and Developmental Biology
ISSN:2296-634X
Year of Completion:2021
Volume:9
Article Number:641618
Source:Frontiers in Cell and Developmental Biology (2021) 9:641618. doi: 10.3389/fcell.2021.641618
DOI:https://doi.org/10.3389/fcell.2021.641618
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Tag:CRISPR-Cas9; JUN; MYC; lung cancer; mouse model; non-small cell lung cancer
Release Date:2022/02/04
Date of first Publication:2021/03/02
Open-Access-Publikationsfonds / Förderzeitraum 2021
Licence (German):License LogoCC BY: Creative-Commons-Lizenz: Namensnennung 4.0 International