4.7 Article

An Influence of Modification with Phosphoryl Guanidine Combined with a 2′-O-Methyl or 2′-Fluoro Group on the Small-Interfering-RNA Effect

Journal

Publisher

MDPI
DOI: 10.3390/ijms22189784

Keywords

modified siRNA; phosphoryl guanidine group; RNA interference

Funding

  1. Russian Science Foundation [19-15-00217]
  2. Russian Science Foundation [19-15-00217] Funding Source: Russian Science Foundation

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The study showed that siRNA modified with the phosphoryl guanidine (PG) group exhibited increased resistance to RNase A but decreased thermodynamic stability. However, PG-modified siRNA retained its activity when the modifications were introduced into the passenger strand.
Small interfering RNA (siRNA) is the most important tool for the manipulation of mRNA expression and needs protection from intracellular nucleases when delivered into the cell. In this work, we examined the effects of siRNA modification with the phosphoryl guanidine (PG) group, which, as shown earlier, makes oligodeoxynucleotides resistant to snake venom phosphodiesterase. We obtained a set of siRNAs containing combined modifications PG/2 '-O-methyl (2 '-OMe) or PG/2 '-fluoro (2 '-F); biophysical and biochemical properties were characterized for each duplex. We used the UV-melting approach to estimate the thermostability of the duplexes and RNAse A degradation assays to determine their stability. The ability to induce silencing was tested in cultured cells stably expressing green fluorescent protein. The introduction of the PG group as a rule decreased the thermodynamic stability of siRNA. At the same time, the siRNAs carrying PG groups showed increased resistance to RNase A. A gene silencing experiment indicated that the PG-modified siRNA retained its activity if the modifications were introduced into the passenger strand.

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