RNA interference (RNAi) is a valuable technique to determine gene function. In Caenorhabditis elegans, RNAi can be achieved by feeding worms bacteria carrying a plasmid expressing double-stranded RNA (dsRNA) targeting a gene of interest. The most commonly used plasmid vector for this purpose is L4440. However, it has been noticed that sequences within L4440 may elicit unspecific effects. Here, we provide a comprehensive characterization of these effects and their mechanisms and describe new unexpected phenotypes uncovered by the administration of unspecific exogenous dsRNA. An example involves dsRNA produced by the multiple cloning site (MCS) of L4440, which shares complementary sequences with some widely used reporter vectors and induces partial transgene silencing via the canonical and antiviral RNAi pathway. Going beyond transgene silencing, we found that the reduced embryonic viability of mir-35-41(gk262) mutants is partially reversed by exogenous dsRNA via a mechanism that involves canonical RNAi. These results indicate cross-regulation between different small RNA pathways in C. elegans to regulate embryonic viability. Recognition of the possible unspecific effects elicited by RNAi vectors is important for rigorous interpretation of results from RNAi-based experiments.

Recent studies have shown that phenolic compounds present in yerba mate have antioxidant defense properties. To verify whether Ilex paraguariensis extracts are capable of increasing the lifespan of an organism, we have used the free-living nematode Caenorhabditis elegans. Notably, this is the first study that analyzes the effects of the extracts of yerba mate obtained from an extraction method that mimics the manner that the plant is consumed by the population by using a live organism. Yerba mate was purchased from commercial markets from Argentina, Brazil, and Uruguay. Ilex paraguariensis extracts significantly increased the life span of C. elegans. Moreover, the extracts reduced the ROS levels per se, and protected from the reduced survival and reproduction rate induced by paraquat exposure. Considering molecular aspects, we observed that the worms pretreated with the extracts depicted higher translocation of the transcription factor DAF-16::GFP to the nucleus. However, there was no increase in the levels of the DAF-16 target genes, SOD-3 and catalase. Our results suggest that the increase of lifespan caused by the different extracts is associated to the antioxidant potential of yerba mate, however this effect is not completely mediated by daf-16.

Salvia hispanica seeds have been commonly used by people that seek healthy habits through natural foods to reduce cholesterol and triacylglycerides levels, however, the evidences that support this assumption are still scarce in literature. Here, we aimed to evaluate the lipid lowering effects of chia by using Caenorhabditis elegans as animal model, a nematode that has proven its usefulness for lipid metabolism studies. We prepared hexane (HE) and Bligh-Dyer (BDE) extracts, evaluated and compared their safety, antioxidant potential and their lipid-lowering activity in the worms. The characterization of both extracts demonstrated that there were no differences in their lipid composition; however, BDE depicted better antioxidant potential. Both extracts reduced worm’s survival from 2%, and reproduction was reduced following treatment with both extracts, though a more notable effect was observed in HE-treated worms. In addition, the non-toxic concentration of both extracts (1%) increased stress resistance against paraquat toxicity in an antidote paradigm. Notably, this same concentration of both extracts reduced lipid accumulation in obese worms, which was not caused by food deprivation. Taken together, our data demonstrate that both extraction methods from chia seeds result in oils that are rich in mono and polyunsaturated fatty acids, which may modulate lipid accumulation and provide antioxidant resistance in C. elegans.