Inhibition of Mycobacterium smegmatis Gene Expression and Growth Using Antisense Peptide Nucleic Acids
- 1 November 2005
- journal article
- Published by S. Karger AG in Microbial Physiology
- Vol. 9 (2) , 101-109
- https://doi.org/10.1159/000088840
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.Keywords
This publication has 43 references indexed in Scilit:
- Peptide Nucleic Acid Antisense Oligomer as a Therapeutic Strategy against Bacterial Infection: Proof of Principle Using Mouse Intraperitoneal InfectionAntimicrobial Agents and Chemotherapy, 2005
- Antimicrobial synergy between mRNA- and protein-level inhibitorsJournal of Antimicrobial Chemotherapy, 2005
- Antisense phosphorodiamidate morpholino oligomer inhibits viability of Escherichia coli in pure culture and in mouse peritonitisJournal of Antimicrobial Chemotherapy, 2005
- Inhibition of Gene Expression inEscherichia coliby Antisense Phosphorodiamidate Morpholino OligomersAntimicrobial Agents and Chemotherapy, 2003
- Inhibition of InhA Activity, but Not KasA Activity, Induces Formation of a KasA-containing Complex in MycobacteriaPublished by Elsevier ,2003
- Essential Genes Are More Evolutionarily Conserved Than Are Nonessential Genes in BacteriaGenome Research, 2002
- Detection of rpoB Mutations in Mycobacterium tuberculosis by Biprobe AnalysisJournal of Clinical Microbiology, 2001
- Peptide nucleic acids as therapeutic agentsCurrent Opinion in Structural Biology, 1999
- PNA, antisense, and antimicrobialsNature Biotechnology, 1998
- Applications for green fluorescent protein (GFP) in the study of hostpathogen interactionsGene, 1996