Fig. 1

CRISPR-mediated generation of IPC and enhanced MSC-derived immunotherapies. Expression of the nuclease-deficient dCas9 fused to a transcriptional activator in MSCs facilitates the activation of endogenous gene expression. To drive the differentiation of MSCs into IPCs (blue boxes), sgRNAs targeting the promoter of target genes such as Pdx-1, NeuroD1, MafA, etc., are delivered to MSCs expressing the dCas9-transcription activator fusion. Additional sgRNAs targeting the promoter of genes involved in MSC immunomodulation (red boxes) are delivered in combination to maintain the immunomodulatory phenotype of MSCs through multiple cell expansions for the development of therapeutic doses of cell therapy. CCL chemokine ligand, CXCR chemokine receptor, dCas9 nuclease-deficient CRISPR-associated protein 9, GCG glucagon, IDO indoleamine 2,3-deoxygenase, IFN interferon, IL interleukin, INS insulin, MafA v-maf musculoaponeurotic fibrosarcoma A, M-CSF macrophage colony stimulating factor, Neurod1 neuronal differentiation 1, Ngn3 neurogenin 3, Nkx6.1 NK6 homeobox 1, NOS nitric oxide synthase, Pdx1 pancreatic and duodenal homeobox 1, sgRNA short guide RNA, SST somatostatin, VEGF vascular endothelial growth factor