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The translation start codon region is sensitive to antisense PNA inhibition in Escherichia coli
KTH, Tidigare Institutioner                               , Bioteknologi.
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2003 (Engelska)Ingår i: Oligonucleotides, ISSN 1545-4576, Vol. 13, nr 6, s. 427-433Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

Antisense peptide nucleic acids (PNA) can inhibit bacterial gene expression with gene and sequence specificity. Using attached carrier peptides that aid cell permeation, the antisense effects when targeting essential genes are sufficient to prevent growth and even kill bacteria. However, many design uncertainties remain, including the difficult question of target sequence selection. In this study, we synthesized 90 antisense peptide-PNAs to target sequences in a head to tail manner across the entire length of the mRNA encoding beta-lactamase. The results from this scan pointed to the start codon region as most sensitive to inhibition. To confirm and refine the result, a higher-resolution scan was conducted over the start codon region of the beta-lactamase gene and the essential Escherichia coli acpP gene. For both genes, the start codon region, including the Shine-Dalgarno motif, was sensitive, whereas antisense agents targeted outside of this region were largely ineffective. These results are in accord with natural antisense mechanisms, which typically hinder the start codon region, and the sensitivity of this region should hold true for most bacterial genes as well as for other RNase H-independent antisense agents that rely on a steric blocking mechanism. Therefore, although other design parameters are also important, the start codon region in E. coli mRNA is the most reliable target site for antisense PNAs.

Ort, förlag, år, upplaga, sidor
2003. Vol. 13, nr 6, s. 427-433
Nyckelord [en]
gene-expression, messenger-rna, bacteria, hybridization, oligomers
Identifikatorer
URN: urn:nbn:se:kth:diva-23250ISI: 000220140500002OAI: oai:DiVA.org:kth-23250DiVA, id: diva2:341948
Anmärkning
QC 20100525Tillgänglig från: 2010-08-10 Skapad: 2010-08-10Bibliografiskt granskad

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Kuttuva, Gunaratna R.
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