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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2009
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| 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 |
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paper:paper_00278424_v106_n32_p13230_SolerBistue2023-06-08T14:54:23Z Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria Soler Bistué, Alfonso Juan de la Cruz Martín, Fernando Ariel Vozza, Nicolás Federico Zorreguieta, Angeles Tolmasky, Marcelo Eduardo 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 Escherichia coli Oligonucleotides Ribonuclease P RNA, Messenger Bacteria (microorganisms) 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. Fil:Soler Bistué, A.J.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Martín, F.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Vozza, N. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Zorreguieta, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Tolmasky, M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2009 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 |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
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 Escherichia coli Oligonucleotides Ribonuclease P RNA, Messenger Bacteria (microorganisms) |
| spellingShingle |
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 Escherichia coli Oligonucleotides Ribonuclease P RNA, Messenger Bacteria (microorganisms) Soler Bistué, Alfonso Juan de la Cruz Martín, Fernando Ariel Vozza, Nicolás Federico Zorreguieta, Angeles Tolmasky, Marcelo Eduardo Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria |
| topic_facet |
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 Escherichia coli Oligonucleotides Ribonuclease P RNA, Messenger Bacteria (microorganisms) |
| description |
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. |
| author |
Soler Bistué, Alfonso Juan de la Cruz Martín, Fernando Ariel Vozza, Nicolás Federico Zorreguieta, Angeles Tolmasky, Marcelo Eduardo |
| author_facet |
Soler Bistué, Alfonso Juan de la Cruz Martín, Fernando Ariel Vozza, Nicolás Federico Zorreguieta, Angeles Tolmasky, Marcelo Eduardo |
| author_sort |
Soler Bistué, Alfonso Juan de la Cruz |
| title |
Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria |
| title_short |
Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria |
| title_full |
Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria |
| title_fullStr |
Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria |
| title_full_unstemmed |
Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria |
| title_sort |
inhibition of aac(6′)-ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria |
| publishDate |
2009 |
| url |
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 |
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| _version_ |
1768541787186528256 |