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

Autores
Soler Bistué, A.J.C.; Martín, F.A.; Vozza, N.; Ha, H.; Joaquín, J.C.; Zorreguieta, A.; Tolmasky, M.E.
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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.
Fuente
Proc. Natl. Acad. Sci. U. S. A. 2009;106(32):13230-13235
Materia
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)
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by/2.5/ar
Repositorio
Biblioteca Digital (UBA-FCEN)
Institución
Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
OAI Identificador
paperaa:paper_00278424_v106_n32_p13230_SolerBistue
Acceder
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oai_identifier_str paperaa:paper_00278424_v106_n32_p13230_SolerBistue
network_acronym_str BDUBAFCEN
repository_id_str 1896
network_name_str Biblioteca Digital (UBA-FCEN)
spelling Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteriaSoler Bistué, A.J.C.Martín, F.A.Vozza, N.Ha, H.Joaquín, J.C.Zorreguieta, A.Tolmasky, M.E.AminoglycosideAntibiotic resistanceAntisenseNucleic acids analogsRNase Paac(6') Ib enzymeamikacinantisense oligonucleotidebacterial DNAbacterial enzymelocked nucleic acidmessenger RNAnucleaseoligoribonucleotideribonucleaseunclassified drugantibiotic resistancearticlebacterial genecontrolled studyenzymatic degradationEscherichia coligene expressiongenetic analysisgrowth inhibitionin vitro studynonhumanpermeabilitypriority journalAcetyltransferasesAmikacinBase SequenceDNADrug Resistance, BacterialEndocytosisEscherichia coliOligonucleotidesRibonuclease PRNA, MessengerBacteria (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.2009info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12110/paper_00278424_v106_n32_p13230_SolerBistueProc. Natl. Acad. Sci. U. S. A. 2009;106(32):13230-13235reponame:Biblioteca Digital (UBA-FCEN)instname:Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturalesinstacron:UBA-FCENenginfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/2.5/ar2025-09-04T09:48:23Zpaperaa:paper_00278424_v106_n32_p13230_SolerBistueInstitucionalhttps://digital.bl.fcen.uba.ar/Universidad públicaNo correspondehttps://digital.bl.fcen.uba.ar/cgi-bin/oaiserver.cgiana@bl.fcen.uba.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:18962025-09-04 09:48:25.049Biblioteca Digital (UBA-FCEN) - Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturalesfalse
dc.title.none.fl_str_mv Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria
title Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria
spellingShingle Inhibition of aac(6′)-Ib-mediated amikacin resistance by nuclease-resistant external guide sequences in bacteria
Soler Bistué, A.J.C.
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)
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
dc.creator.none.fl_str_mv Soler Bistué, A.J.C.
Martín, F.A.
Vozza, N.
Ha, H.
Joaquín, J.C.
Zorreguieta, A.
Tolmasky, M.E.
author Soler Bistué, A.J.C.
author_facet Soler Bistué, A.J.C.
Martín, F.A.
Vozza, N.
Ha, H.
Joaquín, J.C.
Zorreguieta, A.
Tolmasky, M.E.
author_role author
author2 Martín, F.A.
Vozza, N.
Ha, H.
Joaquín, J.C.
Zorreguieta, A.
Tolmasky, M.E.
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv 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)
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)
dc.description.none.fl_txt_mv 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.
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.
publishDate 2009
dc.date.none.fl_str_mv 2009
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/20.500.12110/paper_00278424_v106_n32_p13230_SolerBistue
url http://hdl.handle.net/20.500.12110/paper_00278424_v106_n32_p13230_SolerBistue
dc.language.none.fl_str_mv eng
language eng
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/2.5/ar
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/2.5/ar
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv Proc. Natl. Acad. Sci. U. S. A. 2009;106(32):13230-13235
reponame:Biblioteca Digital (UBA-FCEN)
instname:Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
instacron:UBA-FCEN
reponame_str Biblioteca Digital (UBA-FCEN)
collection Biblioteca Digital (UBA-FCEN)
instname_str Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
instacron_str UBA-FCEN
institution UBA-FCEN
repository.name.fl_str_mv Biblioteca Digital (UBA-FCEN) - Universidad Nacional de Buenos Aires. Facultad de Ciencias Exactas y Naturales
repository.mail.fl_str_mv ana@bl.fcen.uba.ar
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score 12.623145

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