Development of an ObLiGaRe Doxycycline Inducible Cas9 system for pre-clinical cancer drug discovery

• Published in: Nature communications 2020-09, Vol.11 (1), p.4903-16, Article 4903
• Main Authors: Lundin, Anders, Porritt, Michelle J., Jaiswal, Himjyot, Seeliger, Frank, Johansson, Camilla, Bidar, Abdel Wahad, Badertscher, Lukas, Wimberger, Sandra, Davies, Emma J., Hardaker, Elizabeth, Martins, Carla P., James, Emily, Admyre, Therese, Taheri-Ghahfarokhi, Amir, Bradley, Jenna, Schantz, Anna, Alaeimahabadi, Babak, Clausen, Maryam, Xu, Xiufeng, Mayr, Lorenz M., Nitsch, Roberto, Bohlooly-Y, Mohammad, Barry, Simon T., Maresca, Marcello
• Format: Article
• Language: English
• Subjects: 42; 45/23; 45/44; 59/57; 631/208; 631/337; 631/67; 64/60; Animals; Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; Carcinoma, Non-Small-Cell Lung - drug therapy; Carcinoma, Non-Small-Cell Lung - genetics; Cell Line, Tumor; CRISPR-Associated Protein 9 - genetics; CRISPR-Cas Systems - genetics; Doxycycline - pharmacology; Drug Discovery - methods; Drug Screening Assays, Antitumor - methods; Female; Gene Editing - methods; Gene Expression - drug effects; Gene Expression - genetics; Gene Expression Regulation, Neoplastic - drug effects; Genetic Vectors - genetics; HEK293 Cells; High-Throughput Screening Assays - methods; Humanities and Social Sciences; Humans; Lung Neoplasms - drug therapy; Lung Neoplasms - genetics; Male; Mice; Mice, Transgenic; multidisciplinary; Recombination, Genetic - drug effects; Reproducibility of Results; RNA, Guide, CRISPR-Cas Systems - genetics; Science; Science (multidisciplinary); Transcriptional Activation - drug effects; Transfection - methods; Transgenes - genetics
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-18548-9

The CRISPR-Cas9 system has increased the speed and precision of genetic editing in cells and animals. However, model generation for drug development is still expensive and time-consuming, demanding more target flexibility and faster turnaround times with high reproducibility. The generation of a tightly controlled ObLiGaRe doxycycline inducible SpCas9 (ODInCas9) transgene and its use in targeted ObLiGaRe results in functional integration into both human and mouse cells culminating in the generation of the ODInCas9 mouse. Genomic editing can be performed in cells of various tissue origins without any detectable gene editing in the absence of doxycycline. Somatic in vivo editing can model non-small cell lung cancer (NSCLC) adenocarcinomas, enabling treatment studies to validate the efficacy of candidate drugs. The ODInCas9 mouse allows robust and tunable genome editing granting flexibility, speed and uniformity at less cost, leading to high throughput and practical preclinical in vivo therapeutic testing. CRISPR/Cas9 technology has revolutionised the ability of scientists to make genetically modified cells and animal models. Here, the authors make a Tet-On genetically modified mouse using the Streptococcus pyogenes Cas9 and demonstrate that it can be used for generation of mice with lung cancer.