Direct conversion of somatic cells into ‘insulin-producing-cells’ by user-defined multiplex-epigenetic-engineering vector (MEEV-β)

We demonstrate here a single-step and user-friendly approach to generate insulin producing cells by gRNA driven specific-activation of PDX1, NKX6.1, MAFA, Insulin and Glut2 genes in somatic cells via multiplex-epigenetic-engineering-vector (MEEV-β) containing dCas9.P300 core developed by us. Sorted Glut2 + cells could secrete insulin in response to glucose challenge and showed expression of β-cell specific transcription factors: NKX2.2, and aforementioned genes. Expression of Cav1.3, GSK3β,, KJNC11, and SLC30A8 genes substantiated the functional insulin secreting machinery genes in these Glut2 + cells. Also, absence of ARX and GCG expression in these cells highlighted the specificity of the conversion.