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RNA Stability

RNA Stability

Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...

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Sequence-specific and Selective Recognition of Double-stranded RNAs over Single-stranded RNAs by Chemically Modified Peptide Nucleic Acids

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DNA as Tunable Adaptor for siRNA Polyplex Stabilization and Functionalization.

Philipp Heissig¹, Philipp M Klein¹, Philipp Hadwiger²

¹Pharmaceutical Biotechnology, Department of Pharmacy, Center for Nanoscience, Ludwig-Maximilians-Universität, Munich, Germany.

Molecular Therapy. Nucleic Acids

|March 2, 2016

Summary

DNA adaptors enhance the delivery of small interfering RNA (siRNA) for gene silencing. Attaching a DNA oligomer to siRNA significantly improves polyplex stability and transfection efficiency, outperforming multi-siRNA constructs.

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Area of Science:

Biochemistry
Molecular Biology
Nanotechnology

Background:

Small interfering RNA (siRNA) and microRNA are key regulators of gene expression via RNA interference.
Cationic polymers are used for intracellular delivery of nucleic acid therapeutics but face challenges with cargo stability and efficiency.
The small size of siRNA leads to less stable polyplexes with polycations, hindering efficient intracellular delivery.

Purpose of the Study:

To improve the stability and intracellular delivery efficiency of siRNA using DNA oligomers as adaptors.
To investigate the optimal DNA adaptor length and attachment site for enhanced gene silencing.
To explore the utility of DNA adaptors for conjugating additional functional domains to siRNA for targeted delivery.

Main Methods:

Synthesis of sequence-defined cationic polymers via solid-phase chemistry.
Complexation of siRNA with cationic polymers, utilizing DNA oligomers as size and charge enhancers.
Attachment of DNA adaptors to the 3' end of the siRNA passenger strand.
Formation of functionalized polyplexes incorporating endosomolytic peptides and targeting ligands.

Main Results:

Extending siRNA with a 181-nucleotide DNA adaptor maximized gene silencing mediated by cationic polymers.
DNA-assisted siRNA constructs demonstrated superior transfection efficiency compared to multi-siRNA scaffolds.
Attachment of DNA adaptors at the 3' end of the siRNA passenger strand was more effective.
Functionalized polyplexes with DNA adaptors successfully delivered siRNA for targeted gene silencing in folate receptor-positive cells.

Conclusions:

DNA oligomers serve as effective adaptors to enhance siRNA stability and improve cationic polymer-mediated transfection.
A single DNA adaptor of optimal length can significantly boost gene silencing efficacy.
DNA adaptors offer a versatile platform for creating multifunctional siRNA delivery systems for targeted therapeutics.