Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria

Inhibition of bacterial gene expression by RNase P-directed cleavage is a promising strategy for the development of antibiotics and pharmacological agents that prevent expression of antibiotic resistance. The rise in multiresistant bacteria harboring AAC(6′)-Ib has seriously limited the effectivenes...

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Detalles Bibliográficos
Autores principales: Soler Bistué, Alfonso Juan de la Cruz, Martín, Fernando Ariel, Vozza, Nicolás Federico, Zorreguieta, Angeles, Tolmasky, Marcelo Eduardo
Publicado: 2009
Materias:
Aminoglycoside
Antibiotic resistance
Antisense
Nucleic acids analogs
RNase P
aac(6') Ib enzyme
amikacin
antisense oligonucleotide
bacterial DNA
bacterial enzyme
locked nucleic acid
messenger RNA
nuclease
oligoribonucleotide
ribonuclease
unclassified drug
antibiotic resistance
article
bacterial gene
controlled study
enzymatic degradation
Escherichia coli
gene expression
genetic analysis
growth inhibition
in vitro study
nonhuman
permeability
priority journal
Acetyltransferases
Amikacin
Base Sequence
DNA
Drug Resistance, Bacterial
Endocytosis
Oligonucleotides
Ribonuclease P
RNA, Messenger
Bacteria (microorganisms)
Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00278424_v106_n32_p13230_SolerBistue
http://hdl.handle.net/20.500.12110/paper_00278424_v106_n32_p13230_SolerBistue
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
Descripción
Sumario:Inhibition of bacterial gene expression by RNase P-directed cleavage is a promising strategy for the development of antibiotics and pharmacological agents that prevent expression of antibiotic resistance. The rise in multiresistant bacteria harboring AAC(6′)-Ib has seriously limited the effectiveness of amikacin and other aminoglycosides. We have recently shown that recombinant plasmids coding for external guide sequences (EGS), short antisense oligoribonucleotides (ORN) that elicit RNase P-mediated cleavage of a target mRNA, induce inhibition of expression of aac(6′)-Ib and concomitantly induce a significant decrease in the levels of resistance to amikacin. However, since ORN are rapidly degraded by nucleases, development of a viable RNase P-based antisense technology requires the design of nuclease resistant RNA analog EGSs. We have assayed a variety of ORN analogs of which selected LNA/DNA co-oligomers elicited RNase P-mediated cleavage of mRNA in vitro. Although we found an ideal configuration of LNA/DNA residues, there seems not to be a correlation between number of LNA substitutions and level of activity. Exogenous administration of as low as 50 nM of an LNA/DNA co-oligomer to the hyperpermeable E. coli AS19 harboring the aac(6′)-Ib inhibited growth in the presence of amikacin. Our experiments strongly suggest an RNase P-mediated mechanism in the observed antisense effect.

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