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Research Article

Inhibition of Mycobacterium smegmatis Gene Expression and Growth Using Antisense Peptide Nucleic Acids

Kulyté A.a · Nekhotiaeva N.a · Awasthi S.K.b · Good L.a

Author affiliations

aCenter for Genomics and Bioinformatics, Karolinska Institutet, Stockholm, Sweden; bChemical Biology Research Laboratory, Department of Chemistry, University of Delhi, Delhi, India

Related Articles for ""

J Mol Microbiol Biotechnol 2005;9:101–109

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Article / Publication Details

First-Page Preview
Abstract of Research Article

Published online: November 28, 2005
Issue release date: November 2005

Number of Print Pages: 9
Number of Figures: 5
Number of Tables: 2

ISSN: 1464-1801 (Print)
eISSN: 1660-2412 (Online)

For additional information: http://www.karger.com/MMB

Abstract

Antisense agents that inhibit genes at the mRNA level are attractive tools for genome-wide studies and drug target validation. The approach may be particularly well suited to studies of bacteria that are difficult to manipulate with standard genetic tools. Antisense peptide nucleic acids (PNA) with attached carrier peptides can inhibit gene expression in Escherichia coli and Staphylococcus aureus. Here we asked whether peptide-PNAs could mediate antisense effects in Mycobacterium smegmatis. We first targeted the gfp reporter gene and observed dose- and sequence-dependent inhibition at low micromolar concentrations. Sequence alterations within both the PNA and target mRNA sequences eliminated inhibition, strongly supporting an antisense mechanism of inhibition. Also, antisense PNAs with various attached peptides showed improved anti-gfp effects. Two peptide-PNAs targeted to the essential gene inhA were growth inhibitory and caused cell morphology changes that resemble that of InhA-depleted cells. Therefore, antisense peptide-PNAs can efficiently and specifically inhibit both reporter and endogenous essential genes in mycobacteria.

© 2005 S. Karger AG, Basel


Article / Publication Details

First-Page Preview
Abstract of Research Article

Published online: November 28, 2005
Issue release date: November 2005

Number of Print Pages: 9
Number of Figures: 5
Number of Tables: 2

ISSN: 1464-1801 (Print)
eISSN: 1660-2412 (Online)

For additional information: http://www.karger.com/MMB


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