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Evaluation of siRNA Stability and Interaction with Serum Components Using an Agarose Gel-Based Single-Molecule FRET Labeling Method

Methods Mol Biol. 2021-01; 
Martina Tuttolomondo, Henrik J Ditzel
Products/Services Used Details Operation
Custom DNA/RNA Oligos … Fluorescent-labeled oligonucleotides for the sense and antisense siRNA can be obtained from a synthesis service (eg, GenScript) or produced in house (see Note 1). Resuspend the oligonucleotides at 20 μM in RNase-free water. To anneal the two siRNA strands, mix equal … Get A Quote

摘要

Small interfering RNAs (siRNAs) are RNA molecules with promising therapeutic potential as a result of their selective mRNA cleavage. However, despite recent progress, low stability in the bloodstream is an impediment to successful administration in vivo. Thus, the availability of flexible and rapid methods for studying siRNA stability and vehicles is crucial for future novel siRNA-based therapeutics. Herein, we report a fast Förster resonance energy transfer (FRET) method based on agarose gel electrophoresis to evaluate the stability of siRNA in serum as well as siRNA interaction with serum proteins and enzymes.

关键词

Agarose gel electrophoresis, Förster resonance energy transfer, RNA interference, Serum protein corona, Small interfering RNA, siRNA stability