Knockdown of Genes Involved in Transcription and Splicing Reveals Novel RNAi Targets for Pest Control
RNA interference (RNAi) is a promising next generation technology for the development of species-specific pest management. The key to successful RNAi based-plant protection is dependent in part on data-driven target gene selection, a challenging task due to the absence of laboratory strains and the...
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Veröffentlicht in: | Frontiers in agronomy 2021-09, Vol.3 |
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Zusammenfassung: | RNA interference (RNAi) is a promising next generation technology for the development of species-specific pest management. The key to successful RNAi based-plant protection is dependent in part on data-driven target gene selection, a challenging task due to the absence of laboratory strains and the seasonality of most pest species. In this study, we aimed to identify novel target genes by performing a knowledge-based approach in order to expand the spectrum of known potent RNAi targets. Recently, the protein-coding genes
ncm, Rop, RPII-140
, and
dre4
have been identified as sensitive RNAi targets for pest control. Based on these potent RNAi targets, we constructed an interaction network and analyzed a selection of 30 genes in the model beetle
Tribolium castaneum
via injection of dsRNA synthesized by
in vitro
transcription. Nineteen of these targets induced significant mortality of over 70%, including six that caused 100% lethality. Orthologs of active
T. castaneum
RNAi targets were verified in the economically important coleopteran pests
Diabrotica virgifera virgifera
and
Brassicogethes aeneus
. Knockdown of
D. v. virgifera
genes coding for transcription factor
Spt5, Spt6
, and RNA polymerase II subunit RPII-33 caused over 90% mortality in larval feeding assays. Injection of dsRNA constructs targeting RPII-215 or the pre-mRNA-processing factor Prp19 into adult
B. aeneus
resulted in high lethality rates of 93 and 87%, respectively. In summary, the demonstrated knowledge-based approaches increased the probability of identifying novel lethal RNAi target genes from 2% (whole genome) to 36% (transcription- and splicing-related genes). In addition, performing RNAi pre-screening in a model insect increased also the probability of the identification essential genes in the difficult-to-work-with pest species
D. v. virgifera
and
B. aeneus
. |
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ISSN: | 2673-3218 2673-3218 |
DOI: | 10.3389/fagro.2021.715823 |