Quantification of dsRNA using stable isotope labeling dilution liquid chromatography/mass spectrometry

Quantification of dsRNA using stable isotope labeling dilution liquid chromatography/mass... INTRODUCTIONExploitation of the RNA interference (RNAi) pathway to block the expression of specific genes holds considerable promise for the development of novel RNAi‐based insect management strategies. There are a wide range of future potential applications of RNAi to control agricultural insect pests as well as its use for prevention of diseases in beneficial insects. Recent developments in RNAi have created a need for cost‐effective, large‐scale synthesis of double‐stranded RNA (dsRNA), which in turn requires robust analytical techniques to fully characterise and accurately quantify dsRNA prior to RNAi applications. A wide range of dsRNA products can be generated either via bacterial expression systems, in planta or in vitro transcription. The development of suitable analytical methods to characterise the dsRNA products remains a significant challenge.E. coli‐mediated delivery of dsRNA has been reported in C. elegans, planarians, Entamoeba histolytica and Spodoptera exigua. Furthermore, a number of RNAi‐based insect management strategies have also employed the ingestion of bacteria‐expressing dsRNA, application of chemically synthesised dsRNA and transgenic plants expressing dsRNA. To ensure RNAi gene silencing using the above approaches, it is important to both produce and deliver the required amounts of dsRNA. Therefore, the necessary analytical tools to quantify the dsRNA are important to http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Rapid Communications in Mass Spectrometry Wiley

Quantification of dsRNA using stable isotope labeling dilution liquid chromatography/mass spectrometry

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Publisher
Wiley Subscription Services, Inc., A Wiley Company
Copyright
Copyright © 2018 John Wiley & Sons, Ltd.
ISSN
0951-4198
eISSN
1097-0231
D.O.I.
10.1002/rcm.8074
Publisher site
See Article on Publisher Site

Abstract

INTRODUCTIONExploitation of the RNA interference (RNAi) pathway to block the expression of specific genes holds considerable promise for the development of novel RNAi‐based insect management strategies. There are a wide range of future potential applications of RNAi to control agricultural insect pests as well as its use for prevention of diseases in beneficial insects. Recent developments in RNAi have created a need for cost‐effective, large‐scale synthesis of double‐stranded RNA (dsRNA), which in turn requires robust analytical techniques to fully characterise and accurately quantify dsRNA prior to RNAi applications. A wide range of dsRNA products can be generated either via bacterial expression systems, in planta or in vitro transcription. The development of suitable analytical methods to characterise the dsRNA products remains a significant challenge.E. coli‐mediated delivery of dsRNA has been reported in C. elegans, planarians, Entamoeba histolytica and Spodoptera exigua. Furthermore, a number of RNAi‐based insect management strategies have also employed the ingestion of bacteria‐expressing dsRNA, application of chemically synthesised dsRNA and transgenic plants expressing dsRNA. To ensure RNAi gene silencing using the above approaches, it is important to both produce and deliver the required amounts of dsRNA. Therefore, the necessary analytical tools to quantify the dsRNA are important to

Journal

Rapid Communications in Mass SpectrometryWiley

Published: Jan 15, 2018

References

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