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[Modified Oligonucleotides for Guiding RNA Cleavage Using Bacterial RNase P].

D S Novopashina1,2,3, A S Nazarov1,2, M A Vorobjeva1

  • 1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090 Russia.

Molekuliarnaia Biologiia
|January 12, 2019
PubMed
Summary
This summary is machine-generated.

Modified external guide sequences (EGS oligonucleotides) show potential as antibacterial agents by selectively degrading bacterial RNA. These stable EGS oligonucleotides offer a promising approach for targeted RNA hydrolysis.

Keywords:
2'-fluoro modified oligoribonucleotidesEGS oligonucleotidesbacterial RNase Pmodified oligonucleotidesoligo(2'-О-methylribonucleotides)phosphoryl guanidine oligonucleotides

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Antimicrobial Research

Background:

  • Bacterial ribonuclease P (RNase P) is a crucial enzyme for bacterial RNA processing.
  • External guide sequences (EGS oligonucleotides) can direct RNase P to cleave specific RNA targets.
  • Developing stable and effective EGS oligonucleotides for therapeutic applications is an ongoing challenge.

Purpose of the Study:

  • To investigate the efficacy of novel modified external guide sequences (EGS oligonucleotides) in inducing target RNA hydrolysis mediated by bacterial ribonuclease P.
  • To identify modifications that enhance EGS oligonucleotide stability in biological environments while preserving their RNA-cleaving activity.
  • To explore the potential of these modified EGS oligonucleotides as novel antibacterial agents.

Main Methods:

  • Synthesis and characterization of modified EGS oligonucleotides with alterations in the sugar-phosphate backbone.
  • Assessment of the ability of modified EGS oligonucleotides to direct bacterial RNase P for targeted RNA cleavage.
  • Evaluation of the stability of modified EGS oligonucleotides in simulated biological media.
  • Identification of optimal modification patterns for enhanced stability and functional activity.

Main Results:

  • Several modified EGS oligonucleotides demonstrated efficient hydrolysis of a model RNA target using bacterial RNase P.
  • Modifications at the 2'-position (2'-O-methyl and 2'-fluoro) and internucleotide phosphates (phosphoryl guanidines) enhanced stability.
  • Phosphoryl guanidine analogues of oligodeoxyribonucleotides showed stability in biological media and induced RNA hydrolysis.
  • A balance between functional activity and stability was achieved with specific modifications.

Conclusions:

  • Modified EGS oligonucleotides, particularly those with 2'-position or phosphoryl guanidine modifications, are effective in directing bacterial RNase P for targeted RNA cleavage.
  • These modified EGS oligonucleotides exhibit enhanced stability in biological environments, making them promising candidates for therapeutic development.
  • The findings highlight the potential of optimized EGS oligonucleotides as a novel class of antibacterial agents for targeted RNA degradation.