IIBBA   05544
INSTITUTO DE INVESTIGACIONES BIOQUIMICAS DE BUENOS AIRES
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Non-hydrolizable antisense oligonucleotides reduce cell viability in E. coli AS19
Autor/es:
CAROL DAVIES SALA; ANGELES ZORREGUIETA; MARCELO E. TOLMASKY
Lugar:
Rosario
Reunión:
Congreso; IX Congreso Argentino de Microbiología General; 2013
Institución organizadora:
Sociedad Argentina de Microbiología General
Resumen:
The rise on multiresistant bacterial infections is a topic of growing concern, hence the need for novel antimicrobial strategies. External guide sequences (EGS)-technology is a kind of antisense technology that could serve as part of the solution to this problem. EGSs are short antisense oligoribonucleotides that induce RNase P-mediated cleavage of a target RNA. Since oligoribonucleotides are rapidly degraded by nucleases, a practical utilization of EGS technology requires the design of isosequential non-hydrolyzable analogs that mimic the effect of the RNA EGSs. We recently proved that co-oligomers with combinations of locked nucleotides and deoxynucleotides (LNA/DNA) are stable inside and outside bacterial cells and are capable of eliciting RNase P-mediated degradation of a target mRNA. FtsZ, the principal component of the divisome, is the most conserved bacterial cell division gene and is a potential target for new antimicrobials. Our prior work led to the identification of an EGS, called EGSb1, that elicits RNase P degradation of ftsZ mRNA in vitro with high efficiency. To determine if this EGS was active in vivo it was added to cultures of the hyperpermeable E. coli AS19 strain, followed by CFU/ml determination and microscopy examination. Controls were carried out using a sense EGS, EGSb1sense, and another one, EGSAP, that targets the phoA gene. Our results show that there was modest but specific and significant reduction in the CFU/ml in the cultures of cells treated with EGSb1 as compared to those treated with control EGSs, EGSb1sense or EGSAP (p