Abstract
BackgroundRNA interference (RNAi) is a powerful method to inhibit gene expression in a sequence specific manner.ResultsHere, we described the development of RNAi by micro-injection of double-stranded RNA (dsRNA) in the pea aphid Acyrthosiphon pisum. Injection of dsRNA into whole aphid body induced the silencing of two marker genes with different expression patterns: the ubiquitously expressed Ap-crt genes encoding a calreticulin and the gut specific Ap-cath-L gene encoding a cathepsin-L. Time-course analysis of the silencing showed similar temporal patterns for both genes: inhibition started at 1 day after injection, reached its maximum at 5 days and stopped at 7 days. A comparable 40% decrease of gene expression was observed for Ap-crt and Ap-cath-L.ConclusionThe pea aphid is the first Hemipteran insect for which genome sequence will be available soon. The gene knockdown technique developed in this study will be an essential post-genomic tool for further investigations in aphidology.
Highlights
RNA interference (RNAi) is a powerful method to inhibit gene expression in a sequence specific manner
Dicer RNaseIII-type enzymes cleave cytoplasmic double-stranded RNA (dsRNA) into small interfering RNAs duplexes composed of approximately 21 nucleotides. siRNAs duplexes are incorporated into a multiprotein RNA-inducing silencing complex (RISC) where the antisense strand guide RISC to its homologous target mRNA for endonucleolytic cleavage
This work confirms the effectiveness of RNAi in various tissues of the pea aphid and demonstrates that long dsRNA can be substituted to siRNA to induce the inhibition of gene expression in aphids [18]
Summary
RNA interference (RNAi) is a powerful method to inhibit gene expression in a sequence specific manner. SiRNAs duplexes are incorporated into a multiprotein RNA-inducing silencing complex (RISC) where the antisense strand guide RISC to its homologous target mRNA for endonucleolytic cleavage. This mechanism leads to the degradation of the targeted mRNA [1]. Microinjection is widely used in multiple insect species such as mosquitoes, beetles, honey bees and grasshoppers [4,5,7,8,9]. In these insects, RNAi knockdown has been developed for various genes encod-
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