Journal article
Evidence of a tick RNAi pathway by comparative genomics and reverse genetics screen of targets with known loss-of-function phenotypes in Drosophila
BMC Molecular Biology, Vol.10(1)
2009
Abstract
Background
The Arthropods are a diverse group of organisms including Chelicerata (ticks, mites, spiders), Crustacea (crabs, shrimps), and Insecta (flies, mosquitoes, beetles, silkworm). The cattle tick, Rhipicephalus (Boophilus) microplus, is an economically significant ectoparasite of cattle affecting cattle industries world wide. With the availability of sequence reads from the first Chelicerate genome project (the Ixodes scapularis tick) and extensive R. microplus ESTs, we investigated evidence for putative RNAi proteins and studied RNA interference in tick cell cultures and adult female ticks targeting Drosophila homologues with known cell viability phenotype.
Results
We screened 13,643 R. microplus ESTs and I. scapularis genome reads to identify RNAi related proteins in ticks. Our analysis identified 31 RNAi proteins including a putative tick Dicer, RISC associated (Ago-2 and FMRp), RNA dependent RNA polymerase (EGO-1) and 23 homologues implicated in dsRNA uptake and processing. We selected 10 R. microplus ESTs with >80% similarity to D. melanogaster proteins associated with cell viability for RNAi functional screens in both BME26 R. microplus embryonic cells and female ticks in vivo. Only genes associated with proteasomes had an effect on cell viability in vitro. In vivo RNAi showed that 9 genes had significant effects either causing lethality or impairing egg laying.
Conclusion
We have identified key RNAi-related proteins in ticks and along with our loss-of-function studies support a functional RNAi pathway in R. microplus. Our preliminary studies indicate that tick RNAi pathways may differ from that of other Arthropods such as insects.
Details
- Title
- Evidence of a tick RNAi pathway by comparative genomics and reverse genetics screen of targets with known loss-of-function phenotypes in Drosophila
- Authors/Creators
- S. Kurscheid (Author/Creator) - Murdoch UniversityA.E. Lew-Tabor (Author/Creator) - Murdoch UniversityM. Rodriguez Valle (Author/Creator)A.G. Bruyeres (Author/Creator) - Cooperative Research Centre for Beef Genetic TechnologiesV.J Doogan (Author/Creator) - Cooperative Research Centre for Beef Genetic TechnologiesU.G. Munderloh (Author/Creator) - University of MinnesotaF.D. Guerrero (Author/Creator) - Cooperative Research Centre for Beef Genetic TechnologiesR.A. Barrero (Author/Creator) - Murdoch UniversityM.I. Bellgard (Author/Creator) - Murdoch University
- Publication Details
- BMC Molecular Biology, Vol.10(1)
- Publisher
- BioMed Central
- Identifiers
- 991005544117307891
- Copyright
- © 2009 Kurscheid et al
- Murdoch Affiliation
- Centre for Comparative Genomics
- Language
- English
- Resource Type
- Journal article
- Note
- This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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- Collaboration types
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- Citation topics
- 1 Clinical & Life Sciences
- 1.196 Micro & Long Noncoding RNA
- 1.196.1705 RNAi
- Web Of Science research areas
- Biochemistry & Molecular Biology
- ESI research areas
- Molecular Biology & Genetics