Identification of exosome-like nanovesicle (ENV) associated miRNAs in edible plants

MicroRNAs (miRNAs) are small non-coding RNAs between 18-23nts in size which regulate the translation and stability of target mRNAs. miRNAs are present in dietary plants and are conventionally thought to be degraded during the gastrointestinal digestion process. Recent reports suggest that a few dietary microRNAs may exhibit resistance to this process, enter systemic circulation and exert biological effects on animal physiology, currently known as cross-kingdom regulation. However, such horizontal transfer of miRNAs via different kingdoms is highly likely for miRNAs that are present intrinsic extracellular vesicles which increases their bioavailability. These vesicular structures from plants are known as Exosome-like nanovesicles (ENV). ENVs have been isolated from several edible plants. ENV-derived miRNAs are probably more bioavailable and are spontaneously absorbed in intestinal epithelium to suppress target transcripts in human/microbial/viral kingdoms. Such cross-kingdom regulation exhibited by ENV-miRNAs, if properly investigated and validated, may aid in the development of non-toxic and cost-effective therapeutics to treat human diseases. In this line, we purified ENVs from four edible plants (Soy bean, ginger, amla and turmeric). Small RNA population from these ENVs were isolated and profiled through small RNA sequencing to identify ENV-associated miRNAs enriched in each species. Overall design: To identify the miRNAs enriched in edible plant ENVs from Soybean, ginger, amla and turmeric, plant samples were procured from local markets. Each plant material was procured from two different sources and used as biological replicates. ENVs were purified under identical conditions from these plants by differential centrifugation followed by poly ethylene glycol precipitation. After isolation of total RNA from the ENVs, microRNAs were identified by small RNA sequencing.