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Identification of a double-stranded RNA-degrading nuclease influencing both ingestion and injection RNA interference efficiency in the red flour beetle Tribolium castaneum.
- Source :
-
Insect Biochemistry & Molecular Biology . Oct2020, Vol. 125, pN.PAG-N.PAG. 1p. - Publication Year :
- 2020
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Abstract
- RNA interference (RNAi) efficiency dramatically varies among different insects and among administration methods. Numerous studies have revealed that a poor RNAi response is usually associated with a high double-stranded RNA (dsRNA)-degrading activity. Using the red flour beetle Tribolium castaneum , we conducted genome-wide identification of genes encoding dsRNA-degrading nucleases of the DNA/RNA non-specific endonuclease superfamily. To achieve a robust RNAi response in T. castaneum , four dsRNase genes were identified in the genome that seemed to be the potential factors reducing RNAi efficacy. Analysis of biochemical properties revealed that optimal conditions for the dsRNA-degrading activity were alkaline (pH 8.0) in the absence of Mg2+ at 37 °C. The dsRNA-degrading activity was predominantly present in the gut, and via heterologous expression and RNAi experimentation, gut-specific TcdsRNase1 was confirmed as the major nuclease performing dsRNA degradation. After a knockdown of the TcdsRNase1 nuclease activity, RNAi efficiency improved from 38.6% to 58.9% and from 20.9% to 53.9% for injection and ingestion of dsRNA, respectively. Our results contribute to a comprehensive understanding of the mechanisms influencing dsRNA stability and even RNAi efficiency in T. castaneum and point to a good method for improving RNAi efficiency through downregulation of the relevant nuclease activity. Image 1 • We identified four Tribolium castaneum dsRNase genes from the DNA/RNA non-specific endonuclease family. • Optimal biochemical conditions for the dsRNA-degrading activity were found. • TcdsRNase1 was the major nuclease degrading dsRNA. • A knockdown of TcdsRNase1 nuclease activity improved efficiency of RNAi based on either injection or ingestion of dsRNA. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 09651748
- Volume :
- 125
- Database :
- Academic Search Index
- Journal :
- Insect Biochemistry & Molecular Biology
- Publication Type :
- Academic Journal
- Accession number :
- 145739141
- Full Text :
- https://doi.org/10.1016/j.ibmb.2020.103440