AI Article Synopsis
- The study developed a reverse transcription quantitative polymerase chain reaction (RT-qPCR) method to accurately measure chemically modified small interfering RNA (siRNA), including thermally destabilizing modifications like glycol nucleic acid (GNA).
- RT-qPCR proved to be more sensitive and capable of higher throughput than mass spectrometry for siRNA detection, though mass spectrometry is preferred for specific metabolite detection due to its inefficacy with certain chemical modifications.
- Both RT-qPCR and mass spectrometry have unique advantages and disadvantages, so the choice of method should depend on factors like throughput, sensitivity, and the need for specific metabolite identification.
Article Abstract
The goal of this study was to develop a reverse transcription quantitative polymerase chain reaction (RT-qPCR) method for the accurate quantification of chemically modified small interfering RNA (siRNA) including but not restricted to thermally destabilizing modifications such as glycol nucleic acid (GNA). RT-qPCR was found to be superior to mass spectrometry-based siRNA detection in terms of sensitivity and throughput. However, mass spectrometry is still the preferred method when specific metabolite detection is required and is also insensitive to siRNA chemical modifications such as GNA. The RT-qPCR approach can be optimized to take chemical modifications into account and works robustly in different matrices without optimization, unlike mass spectrometry. RT-qPCR and mass spectrometry both have their strengths and weaknesses for the detection of siRNA and must be used appropriately depending on the questions at hand. Considerations such as desired throughput, assay sensitivity, and metabolite identification must be weighed when choosing which methodology to apply.
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| http://dx.doi.org/10.1089/nat.2019.0840 | DOI Listing |
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