Modulación de La expresión génica De TNPO3 en distrofia muscular de cintura Lgmd1f

Abstract

Abstract: Limb-girdle muscular dystrophies (LGMD) are heterogeneous genetic diseases that present a great clinical appearance. Within this research work we will focus on the autosomal dominant type LGMDD2, related by the mutation of a nucleotide in the stop codon of the Transportin 3 gene (TNPO3), responsible for the extension of 15 amino acids, which results in the mutant protein (TNPO3 mut), causing phenotypic malformations at the muscle level, both distal and proximal. For this reason, the present research work aims to characterize a cellular model of LGMDD2 and to establish proofs of concept for the use of splice-modified antisense oligonucleotides (ASO) as a possible gene therapy. To this end, ASOs directed at intron 22 and exon 23 of the TNPO3 gene have been designed and the processes for modulating gene expression have been standardized to reduce the synthesis of TNPO3 mut. Hela cells were taken as biological models and the validation of the ASO treatment was quantified by RT- qPCR. A total of 5 ASOs specific for the TNPO3 gene were obtained. It was concluded that these synthetic oligos directed to specific sequences can modulate the expression of the gene, being the present investigation a starting study for the search of possible therapeutic targets.