32 Cytokine-STAT5 Signaling Controls Intestinal Stem Cell Activation to Suppress Clostridium Difficile - Induced Gut Inflammation

injection of guide RNA against Apc (U6-sgApc-CMV-turboRFP) into LSL-Cas9-eGFP-Villin creER mice (treated with tamoxifen) results in epithelial-specific tumor initiation.Figure 2: In situ engraftment of CRISPR/Cas9-mediated Apc-null intestinal organoid bodies results in adenoma formation and sequential mutagenesis.a. Apc editing in wildtype murine intestinal organoids using CRISPR lentiviral infection (U6-sgApc-EFS-Cas9-2A-GFP) results in cystic organoids that exhibit robust Wnt pathway activation.b.Orthotopic transplantation of Apc-null organoids into wild-type recipient mice results in efficient tumor formation.c.CRISPR-mediated Apc deletion in LSL-Kras G12D -Lgr5 creER -LSL-TdTomato colon organoids, followed by orthotopic transplantation, results in tumor formation.Tamoxifen injection activates mutant Kras in a clonal population of established tumor-initiating cells.