![Evolution Mk 149 Drivers](https://kumkoniak.com/78.jpg)
Replacement of CTP/UTP with 5-Hydroxymethyl CTP and UTP Replacement of CTP or UTP with 5-Hydroxymethyl CTP or UTP Replacement of CTP/UTP with 2’Fluoro CTP and UTP Replacement of CTP or UTP with 2’Fluoro CTP or UTP
![Evolution Mk 149 Drivers Evolution Mk 149 Drivers](https://am76.pl/plik/201703211612_3D_miasta.181215.800x0.jpg)
Replacement of NTPs with combinations of α-thiophosphates ATP/CTP/GTP/UTP Phosphorothioate (A) Phosphorothioate (C) Replacement of NTPs with α-thiophosphate ATP or CTP or GTP or UTP Table 1 Composition and nomenclature of chemically modified dsRNA synthesised and used in this study. Finally, the RNAi efficacy of long chemically modified dsRNA was examined both in vitro in Drosophila cell culture using a dual luciferase reporter assay, and in vivo in SGSB nymphs and western corn rootworm ( Diabrotica virgifera virgifera) (WCR) larvae using survival studies. The ability of model RNase III/Dicer family enzymes to successfully cleave long chemically modified dsRNA into endoribonuclease-prepared siRNAs (esiRNAs) in vitro was also investigated. The effects of chemical modifications on the nuclease stability of long dsRNA were studied in vitro using southern green stink bug ( Nezara viridula) (SGSB) saliva, Colorado potato beetle ( Leptinotarsa decemlineata) (CPB) gut secretions, and agricultural soil as examples of sources of nucleases likely to contribute to degradation of insecticidal dsRNA. In this study, we have optimised the synthesis and purification of long dsRNA containing a range of different chemical modifications including phosphorothioate (PS), 2’-fluoro (2’F), and 5-hydroxymethyl (HMr) modifications (see Table 1). These results provide further mechanistic insight into the dependence of RNAi efficacy on nucleotide modifications in the sense or antisense strand of the dsRNA in insects and demonstrate for the first time that RNAi can successfully be triggered by chemically-modified long dsRNAs in insect cells or live insects. In live insects, we found chemically-modified long dsRNAs successfully resulted in mortality in both stink bug and corn rootworm. Phosphorothioate- and 2’-fluoro-modified dsRNA also demonstrated increased resistance to degradation by soil nucleases and increased RNAi efficacy in Drosophila melanogaster cell cultures. Our results showed that dsRNAs containing phosphorothioate modifications demonstrated increased resistance to southern green stink bug saliva nucleases. In this study, we synthesised a range of chemically-modified long dsRNAs in an approach to improve nuclease resistance and RNAi efficacy in insects. However, there are significant challenges associated with RNAi efficacy in insects.
![Evolution Mk 149 Drivers Evolution Mk 149 Drivers](http://www.toyking.com.tw/image/toy/bandai/monster/196425.jpg)
![Evolution Mk 149 Drivers Evolution Mk 149 Drivers](https://am76.pl/plik/20200713212338_6-figurka-do-gry-przygodowej-rpg.jpg)
Long dsRNAs are capable of inducing RNA interference (RNAi) in insects and are emerging as novel, highly selective alternatives for sustainable insect management strategies. Many of these crop pests either are not currently controlled by artificial means or have developed resistance against chemical pesticides. Global agriculture loses over $100 billion of produce annually to crop pests such as insects. Glycobiology and Extracellular Matrices.
![Evolution Mk 149 Drivers](https://kumkoniak.com/78.jpg)