Use of different CRISPR activation systems to activate human beta pancreatic genes

GIMENEZ CARLA; HYON, SUNG HO; CURTI LUCIA; PEREYRA-BONNET FEDERICO

CAPITAL FEDERAL

Congreso; CONGRESO SAIC; 2017

Sociedad Argentina de Investigación Clínica

The CRISPR systems for gene activation are new molecular tools with a promising future in the area of cell reprogramming. However, its convenient use must be explored in order to maximize the efficiency/cost ratio. In this work, we used the CRISPR/dCas9-VP160, CRISPR/dCas9-TET1 and CRISPR/dCas9-P300 systems for the activation of human pancreatic genes in order to emulate a reprogramming protocol towards the pancreatic beta cell. For this, we studied several aspects such as the activation efficiency dependent on the system, quantity and effectiveness of designed sgRNAs, the individual or combined use of functional sgRNAs, the performance in the multiplex activation of genes of the same lineage, the strategy of using a sequential or multiple activations and the possibility and effectiveness of the mRNA delivery of the system. In conclusion, this work proposes a deep exploration of these systems, especially for CRISPR/dCas9-P300, and answers unknowns so far not studied that could serve as a basis to optimize future trials using this molecular tool.