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bioRxiv · 10.1101/2022.08.12.503816

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

Naqvi, R.·Naqvi, A.·Priyadarshini, M.
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Abstract

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-{beta}) containing dCas9.P300core developed by us. Sorted Glut2+ cells could secrete insulin in response to glucose challenge and showed expression of {beta}-cell specific transcription factors: NKX2.2, and aforementioned genes. Expression of Cav1.3, GSK3{beta},, 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.

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Naqvi, R., Naqvi, A., Priyadarshini, M.. 2022-08-13. Direct conversion of somatic cells into insulin-producing-cells by user-defined multiplex-epigenetic-engineering vector (MEEV-b). https://doi.org/10.1101/2022.08.12.503816

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