Attempts to establish RNA interference in the parasitic nematode Heligmosomoides polygyrus

Lendner, Matthias; Doligalska, Maria; Lucius, Richard; Hartmann, Susanne

doi:10.1016/j.molbiopara.2008.06.003

Cited by 42 publications

(53 citation statements)

References 59 publications

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“…The only species of animal-parasitic nematode in which this method has been documented to have a phenotypic effect is Trichostrongylus colubriformis (Issa et al 2005). In the case of H. polygyrus, it was noted that L1 ingest many fewer bacteria than the corresponding stage of C. elegans (Lendner et al 2008). In an attempt to increase the liberation of dsRNA from bacteria in the parasite intestine, we transfected bacteria with pLysE, a plasmid expressing bacteriophage T7 lysozyme, but this proved to be toxic and inhibited larval development (unpublished data).…”

Section: Ingestion Of Bacterially Expressed Dsrnamentioning

confidence: 99%

The development of RNA interference (RNAi) in gastrointestinal nematodes

et al. 2012

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Despite the utility of RNAi for defining gene function in Caenorhabditis elegans and early successes reported in parasitic nematodes, RNAi has proven to be stubbornly inconsistent or ineffective in the animal parasitic nematodes examined to date. Here, we summarise some of our experiences with RNAi in parasitic nematodes affecting animals and discuss the available data in the context of our own unpublished work, taking account of mode of delivery, larval activation, site of gene transcription and the presence/absence of essential RNAi pathway genes as defined by comparisons to C. elegans. We discuss future directions briefly including the evaluation of nanoparticles as a means to enhance delivery of interfering RNA to the target worm tissue.

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Section: Ingestion Of Bacterially Expressed Dsrnamentioning

confidence: 99%

The development of RNA interference (RNAi) in gastrointestinal nematodes

et al. 2012

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“…In several previous studies, cross-species of RNAi has been used to investigate the gene function (Gao et al 2006;Lendner et al 2008), but this approach has not been applied in Trichinella. Although the entire genomic sequencing of Trichinella has been finished, a lot of gene function remains unclear.…”

Section: Discussionmentioning

confidence: 99%

Expression and functional characterization of a Rho-family small GTPase CDC42 from Trichinella spiralis

et al. 2010

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A full-length cDNA encoding a Rho-family small GTPase gene cdc42 of Trichinella spiralis was expressed in E. coli. The recombinant protein of TsCDC42 was purified and used to raise the polyclonal antibodies. The expression of TsCDC42 in different stages of parasite was investigated. The western blot showed that TsCDC42 was expressed in all stages of T. spiralis, including newborn larvae, muscle larvae and adult worms. The immuno-localization revealed that TsCDC42 was ubiquitously distributed in the newborn larvae, muscle larvae and adult worm. Cross-species RNAi was done by knockdown Tscdc42 RNAi in C. elegans. The results revealed that endogenous expression level of CDC42 was decreased by knockdown Tscdc42 RNAi in C. elegans, and this knockdown reduced the progeny of C. elegans. It suggested that Tscdc42 might play the same roles in the early development of T. spiralis.

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“…elegans is often inefficient or inconsistent. In Heligmosomoides polygyrus , RNAi by feeding did not result in phenotypical changes or in significant down-regulation of overall mRNA levels [13]. Similar difficulties have been observed in Haemonchus contortus [14].…”

Section: Introductionmentioning

confidence: 87%

“…RNAi by feeding is a convenient reverse genetic technique that allows nematodes to develop under nearly natural conditions on a bacterial lawn [13]. The use of RNAi in nematode species other than C .…”

Section: Introductionmentioning

confidence: 99%

Establishing RNAi in a Non-Model Organism: The Antarctic Nematode Panagrolaimus sp. DAW1

et al. 2016

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The Antarctic nematode Panagrolaimus sp. DAW1 is one of the only organisms known to survive extensive intracellular freezing throughout its tissues. Although the physiological mechanisms of this extreme adaptation are partly understood, the molecular mechanisms remain largely unknown. RNAi is a method that allows the examination of gene function in a direct, targeted manner, by knocking out specific mRNAs and revealing the effects on the phenotype. In this study we have explored the viability of RNAi in Panagrolaimus sp. DAW1. In the first trial, nematodes were fed E. coli expressing Panagrolaimus sp. DAW1 dsRNA of the embryonic lethal genes rps-2 and dhc, and the blister gene duox. Pd-rps-2(RNAi)-treated nematodes showed a significant decrease in larval hatching. However, qPCR showed no significant decrease in the amount of rps-2 mRNA in Pd-rps-2(RNAi)-treated animals. Several soaking protocols for dsRNA uptake were investigated using the fluorescent dye FITC. Desiccation-enhanced soaking showed the strongest uptake of FITC and resulted in a significant and consistent decrease of mRNA levels of two of the four tested genes (rps-2 and tps-2a), suggesting effective uptake of dsRNA-containing solution by the nematode. These findings suggest that RNAi by desiccation-enhanced soaking is viable in Panagrolaimus sp. DAW1 and provide the first functional genomic approach to investigate freezing tolerance in this non-model organism. RNAi, in conjunction with qPCR, can be used to screen for candidate genes involved in intracellular freezing tolerance in Panagrolaimus sp. DAW1.

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Attempts to establish RNA interference in the parasitic nematode Heligmosomoides polygyrus

Cited by 42 publications

References 59 publications

The development of RNA interference (RNAi) in gastrointestinal nematodes

The development of RNA interference (RNAi) in gastrointestinal nematodes

Expression and functional characterization of a Rho-family small GTPase CDC42 from Trichinella spiralis

Establishing RNAi in a Non-Model Organism: The Antarctic Nematode Panagrolaimus sp. DAW1

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