Fig. 4: ODInCas9 mouse non-small cell lung cancer models.
From: Development of an ObLiGaRe Doxycycline Inducible Cas9 system for pre-clinical cancer drug discovery
a Schematic representation of generation of ODInCas9 KrasG12D;Trp53−/−, KrasG12D;Stk11−/−, and KrasG12C;Trp53−/− non-small cell lung cancer models. ODInCas9 mice are treated with doxycycline 3 days before administration via oral aspiration of model-specific AAV targeting either Kras by sgRNA and HDR and Trp53 or Stk11 by sgRNA. Doxycycline treatment is kept 1 day post viral administration. Lung adenocarcinoma kinetics is specific for each lung cancer model. b Representative HE staining of lung histology capturing the various stages of lung adenocarcinoma. c Western blots showing Trp53 and Stk11 expression in cell lines generated from either Kras;Trp53 or Kras;Stk11 lung tumors and d in multiple independent lung tumors from the Kras;Trp53 and Kras;Stk11 models. MC38, M2, HEK293 cell lines and non-tumor bearing lung shown as positive control. e Classification of amplicon sequencing of lung cancer tissue at Kras and Trp53 locus. f DNA in situ hybridization targeting KrasG12D. g Immunohistochemistry section of lung tumors staining for Ki67, pERK, pMEK, CD31, αSMA, F480, CD45R, CD4/8, pSPC, ɣH2AX, Tenascin C, and Nkx2.1.
