ArticleParasites & vectors2012
Efficient in vitro RNA interference and immunofluorescence-based phenotype analysis in a human parasitic nematode, Brugia malayi.
Article in Parasites & vectors, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed, 34 citations in OpenAlex.
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- Toward novel treatment against filariasis: Insight into genome-wide co-evolutionary analysis of filarial nematodes andFrontiers in microbiology · 2023Article
- Localization and RNA Interference-Driven Inhibition of a Brugia malayi-Encoded Interleukin-5 Receptor Binding Protein.Infection and immunity · 2022Article
- A qPCR to quantify Wolbachia from few Onchocerca volvulus microfilariae as a surrogate for adult worm histology in clinical trials of antiwolbachial drugs.Parasitology research · 2022Article
- The Functional Parasitic Worm Secretome: Mapping the Place ofPathogens (Basel, Switzerland) · 2020Review
- The endosymbiont Wolbachia rebounds following antibiotic treatment.PLoS pathogens · 2020Article
- Terror in the dirt: Sensory determinants of host seeking in soil-transmitted mammalian-parasitic nematodes.International journal for parasitology. Drugs and drug resistance · 2018Review
- Tackling Hypotheticals in Helminth Genomes.Trends in parasitology · 2018Article
- Functional genomics inProceedings of the National Academy of Sciences of the United States of America · 2017Article
- Direct experimental manipulation of intestinal cells in Ascaris suum, with minor influences on the global transcriptome.International journal for parasitology · 2017Article
- Heme acquisition in the parasitic filarial nematode Brugia malayi.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2016Article
- Prospects and challenges of CRISPR/Cas genome editing for the study and control of neglected vector-borne nematode diseases.The FEBS journal · 2016Review
- Glucose and Glycogen Metabolism in Brugia malayi Is Associated with Wolbachia Symbiont Fitness.PloS one · 2016Article
- Rendering the Intractable More Tractable: Tools from Caenorhabditis elegans Ripe for Import into Parasitic Nematodes.Genetics · 2015Review
- A novel member of the let-7 microRNA family is associated with developmental transitions in filarial nematode parasites.BMC genomics · 2015Article
- Genome mining offers a new starting point for parasitology research.Parasitology research · 2015Review
- Comparative analysis of the secretome from a model filarial nematode (Litomosoides sigmodontis) reveals maximal diversity in gravid female parasites.Molecular & cellular proteomics : MCP · 2014Article
- A target repurposing approach identifies N-myristoyltransferase as a new candidate drug target in filarial nematodes.PLoS neglected tropical diseases · 2014Article
- Co-evolution between an endosymbiont and its nematode host: Wolbachia asymmetric posterior localization and AP polarity establishment.PLoS neglected tropical diseases · 2014Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundRNA interference (RNAi) is an efficient reverse genetics technique for investigating gene function in eukaryotes. The method has been widely used in model organisms, such as the free-living nematode Caenorhabditis elegans, where it has been deployed in genome-wide high throughput screens to identify genes involved in many cellular and developmental processes. However, RNAi techniques have not translated efficiently to animal parasitic nematodes that afflict humans, livestock and companion animals across the globe, creating a dependency on data tentatively inferred from C. elegans.
resultsWe report improved and effective in vitro RNAi procedures we have developed using heterogeneous short interfering RNA (hsiRNA) mixtures that when coupled with optimized immunostaining techniques yield detailed analysis of cytological defects in the human parasitic nematode, Brugia malayi. The cellular disorganization observed in B. malayi embryos following RNAi targeting the genes encoding γ-tubulin, and the polarity determinant protein, PAR-1, faithfully phenocopy the known defects associated with gene silencing of their C. elegans orthologs. Targeting the B. malayi cell junction protein, AJM-1 gave a similar but more severe phenotype than that observed in C. elegans. Cellular phenotypes induced by our in vitro RNAi procedure can be observed by immunofluorescence in as little as one week.
conclusionsWe observed cytological defects following RNAi targeting all seven B. malayi transcripts tested and the phenotypes mirror those documented for orthologous genes in the model organism C. elegans. This highlights the reliability, effectiveness and specificity of our RNAi and immunostaining procedures. We anticipate that these techniques will be widely applicable to other important animal parasitic nematodes, which have hitherto been mostly refractory to such genetic analysis.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.