Antisense Effects of PNAs in Bacteria
Antisense peptide nucleic acid (PNA) can be used to control bacterial gene expression. PNAs are designed to target sequences within messenger RNA and knock-down gene expression. PNAs targeted to the translation initiation region of mRNA are particularly effective and result in mRNA degradation and reduced protein expression from the targeted gene. The antisense effects can be sufficient to alter phenotypes and even kill bacteria. PNAs provide useful tools for the study of bacterial gene function, and with improvements in cell uptake antisense PNAs may find applications as antimicrobial agents. This chapter details methods for designing antisense PNAs and testing their activities in bacteria, including suggestions for control experiments.
Key wordsPeptide nucleic acid (PNA) DNA analogue Solid-phase peptide synthesis (SPPS) Metal complexes Click chemistry
- 4.Harth G, Horwitz MA, Tabatadze D, Zamecnik PC (2002) Targeting the Mycobacterium tuberculosis 30/32-kDa mycolyl transferase complex as a therapeutic strategy against tuberculosis: proof of principle by using antisense technology. Proc Natl Acad Sci U S A 99(24):15614–15619PubMedCentralPubMedCrossRefGoogle Scholar
- 25.Dryselius R, Nekhotiaeva N, Good L (2005) Antimicrobial synergy between mRNA-and protein-level inhibitors. Br Soc Antimicrob ChemotherGoogle Scholar
- 28.Gião MS, Wilks SA, Azevedo NF, Vieira MJ, Keevil CW (2009) Comparison between standard culture and peptide nucleic acid 16S rRNA hybridization quantification to study the influence of physico-chemical parameters on Legionella pneumophila survival in drinking water biofilms. Biofouling 25(4):343–351PubMedGoogle Scholar