Issue 5, 2008
From the journal:

Molecular BioSystems

Photoinduced RNA interference using DMNPE-caged 2′-deoxy-2′-fluoro substituted nucleic acidsin vitro and in vivo†

Richard A. Blidner,a   Kurt R. Svoboda,b   Robert P. Hammerc  and  W. Todd Monroe*a  

* Corresponding authors

a Rm. 149 E.B. Doran Bldg., Biological & Agricultural Engineering, Louisiana State University and LSU Agricultural Center, Baton Rouge, USA
E-mail: tmonroe@LSU.edu
Fax: +1-225-578-3492
Tel: +1-225-578-1059

b Department of Biological Sciences, Louisiana State University, Baton Rouge, USA

c Department of Chemistry, Louisiana State University, Baton Rouge, USA

Abstract

Various chemical modifications to RNA have been incorporated in attempts to improve their pharmacological properties for RNAi interference (RNAi). Recent studies have shown that small interfering RNA (siRNA) containing 2′-fluoro modifications can elicit gene silencing through RNAi. Despite developments in using chemical modifications for increased stability, safety, and efficiency of these therapeutics, they still face challenges of spatial and temporal targeting. One potential targeting strategy is to use photocaging techniques, which involve the covalent attachment of photolabile compounds to the effectornucleic acid species that block bioactivity until exposed to near UV light. In this study we demonstrate that fully 2′-fluorinated nucleic acids (FNAs) can be caged for photoactivated gene silencing in cell culture and in zebrafish embryos. This strategy combines the improvement in chemical and enzymatic stability associated with 2′-substitutions with the targeting ability of a photoinducible trigger. Statistical alkylation of FNAs with 1-(4,5-dimethoxy-2-nitrophenyl)diazoethane (DMNPE) improved resistance to enzymatic degradation, reduced RNAi effectiveness, and protected the biological system from toxic doses of the effector. Photo-exposure to 365 nm light partially restored the silencing activity of the 2′-fluoro siRNAs. These results suggest that photocaging may offer control over RNAi therapeutics for spatially and temporally directed activation, while improving enzymatic stability and potentially enabling therapeutic dosing via light dose intensity.

Graphical abstract: Photoinduced RNA interference using DMNPE-caged 2′-deoxy-2′-fluoro substituted nucleic acidsin vitro and in vivo

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Article information

DOI
https://doi.org/10.1039/B801532E
Article type
Paper
Submitted
29 Jan 2008
Accepted
06 Mar 2008
First published
31 Mar 2008

Mol. BioSyst., 2008,4, 431-440

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Photoinduced RNA interference using DMNPE-caged 2′-deoxy-2′-fluoro substituted nucleic acidsin vitro and in vivo

R. A. Blidner, K. R. Svoboda, R. P. Hammer and W. T. Monroe, Mol. BioSyst., 2008, 4, 431 DOI: 10.1039/B801532E

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