Nature Biotechnology
19, 360 - 364 (2001)
doi:10.1038/86753
Bactericidal antisense effects of peptide−PNA conjugatesLiam Good1, Satish Kumar Awasthi2, Rikard Dryselius1, Ola Larsson1
& Peter E. Nielsen2, 31
Center for Genomics Research, Karolinska Institute, Berzelius väg 37, 171 77, Stockholm, Sweden. 2
Center for Biomolecular Recognition, IMBG, Department of Biochemistry B, The Panum Institute, University of Copenhagen, Blegdamsvej 3c, 2200 Copenhagen N., Denmark. 3
Pantheco A/S, Danish Science Park, Bøge Allé 3, DK 2970 Hørsholm, Denmark.
Correspondence should be addressed to Liam Good liam.good@cgr.ki.se or Peter E. Nielsen pen@imbg.ku.dkAntisense peptide nucleic acids (PNAs) can specifically inhibit Escherichia coli gene expression and growth and hold promise as anti-infective agents and as tools for microbial functional genomics. Here we demonstrate that chemical modification improves the potency of standard PNAs. We show that 9- to 12-mer PNAs, especially when attached to the cell wall/membrane-active peptide KFFKFFKFFK, provide improvements in antisense potency in E. coli amounting to two orders of magnitude while retaining target specificity. Peptide−PNA conjugates targeted to ribosomal RNA (rRNA) and to messenger RNA (mRNA) encoding the essential fatty acid biosynthesis protein Acp prevented cell growth. The anti-acpP PNA at 2  M concentration cured HeLa cell cultures noninvasively infected with E. coli K12 without any apparent toxicity to the human cells. These results indicate that peptides can be used to carry antisense PNA agents into bacteria. Such peptide−PNA conjugates open exciting possibilities for anti-infective drug development and provide new tools for microbial genetics.
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