GPP SGRNA DESIGNER VS CHOP CHOP: IMPLICATIONS OF THE GENOMIC CONTEXT IN CRISPRA SGRNA DESIGN FOR ANGIOGENIC GENES

BELAICH MARIANO NICOLÁS; OLEA FD; PERALTA TOMÁS; CROTTOGINI AJ; NÚÑEZ PEDROZO CRISTIAN NAHUEL; GIMENEZ CS; CUNIBERTI L

Buenos Aires

Congreso; Reunión Anual de Sociedades de Biociencia 2020; 2020

Sociedad Argentina de Investigación Clínica y de Fisiología

Introduction: Angiogenic gene overexpression has been the main strategy in numerous cardiovascular regenerative gene therapy projects. However, most have failed in clinical trials, due to, among other reasons, dose inadequacy and lack of potency. CRISPRa technology enhances gene overexpression levels based on the identification of sgRNAs with maximum efficiency and safety. GPP sgRNA Designer and CHOP CHOP are the most widely used platforms for the prediction of sgRNAs, the scope of efficiency and sensitivity of their algorithms being partially uncertain. The objective of our study was to analyze the performance of GPP sgRNA Designer and CHOP CHOP for the design of sgRNAs in a panel of angiogenic genes, in a changing genetic context such as successive versions of the human genome.Materials and methods: The top 20 ranked sgRNAs were provided by GPP sgRNA Designer (GPP) and CHOP CHOP (CHOP) for each of the following VEGFA, KDR, EPO, HIF-1A, HGF, EGF, PGF, FGF1 genes from both GRCH 37 and GRCH 38 human genomes.Results: The mean ranking variation in the 20 positions was greater for GPP than CHOP in EPO (p