Development of an ObLiGaRe Doxycycline Inducible Cas9 system for pre-clinical cancer drug discovery
Anders Lundin, Michelle J. Porritt, Himjyot Jaiswal, Frank Seeliger, Camilla Johansson, Abdel Wahad Bidar, Lukas Badertscher, Sandra Wimberger, Emma Jane Davies, Elizabeth Hardaker, Carla P. Martins, Emily James, Therése Admyre, Amir Taheri‐Ghahfarokhi, Jenna Bradley, Anna Schantz, Babak Alaei-Mahabadi, Maryam Clausen, Xiufeng Xu, Lorenz M. Mayr, Roberto Nitsch, Mohammad Bohlooly‐Y, Simon T. Barry, Marcello Maresca
Abstract
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.