Attention:The NSF Public Access Repository (PAR) system and access will be unavailable from 11:00 PM ET on Thursday, August 13 until 12:00 AM ET on Friday, August 14 due to maintenance. We apologize for the inconvenience.


Title: The Small RNA Universe of Capitella teleta
RNAi is an evolutionarily fluid mechanism with dramatically different activities across animal phyla. One major group where there has been little investigation is annelid worms. Here, the small RNAs of the polychaete developmental model Capitella teleta are profiled across development. As is seen with nearly all animals, nearly 200 microRNAs were found with 58 high-confidence novel species. Greater miRNA diversity was associated with later stages consistent with differentiation of tissues. Outside miRNA, a distinct composition of other small RNA pathways was found. Unlike many invertebrates, an endogenous siRNA pathway was not observed, indicating pathway loss relative to basal planarians. No processively generated siRNA-class RNAs could be found arising from dsRNA precursors. This has a significant impact on RNAi technology development for this group of animals. Unlike the apparent absence of siRNAs, a significant population of piRNAs was observed. For many piRNAs, phasing and ping-pong biogenesis pathways were identified. Interestingly, piRNAs were found to be highly expressed during early development, suggesting a potential role in regulation in metamorphosis. Critically, the configuration of RNAi factors in C. teleta is found in other annelids and mollusks, suggesting that similar biology is likely to be present in the wider clade. This study is the first in providing comprehensive analysis of small RNAs in annelids.  more » « less
Award ID(s):
1845978
PAR ID:
10322001
Author(s) / Creator(s):
; ; ;
Date Published:
Journal Name:
Frontiers in Molecular Biosciences
Volume:
9
ISSN:
2296-889X
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Abstract Mollusks are an abundant group of animals, with many ecologically and economically important members that are phylogenetically distinct from nearly all genetic model organisms. This study provides a clade-wide evaluation of small RNA biogenesis pathways, with emphasis on the eastern oyster, Crassostrea virginica. A more thorough characterization of these molecules supports rationale design of RNA interference (RNAi) approaches for manipulation of mollusk genetics. Like other animal groups, mollusks have conserved microRNAs (miRNAs), with some shared with ecdysozoans and deuterostomes; however, there was no evidence of a dedicated endogenous small-interfering RNA (siRNA) pathway. These findings suggest that alternatives to long double-stranded RNA (dsRNA)-mediated knockdown, specifically short-hairpin RNAs or small duplex RNAs, are likely more appropriate for gene silencing in mollusks. The study also finds abundant Piwi-interacting RNAs (piRNAs) in both soma and gonads with some mollusk-specific aspects. Many invertebrates exhibit somatic piRNAs; however, mollusk piRNAs appear to be restricted to a subset of cells, suggesting that the potential of piRNA-based RNAi is also limited. Further, individual animals also express a unique collection of piRNAs that seem to be only partially determined through inheritance from parents. Together, this work defines the RNAi mechanisms in mollusks and provides insights into the phenotypic diversity seen in this group. 
    more » « less
  2. null (Ed.)
    RNAi promises to reshape pest control by being nontoxic, biodegradable, and species specific. However, due to the plastic nature of RNAi, there is a significant variability in responses. In this study, we investigate small RNA pathways and processing of ingested RNAi trigger molecules in a hemipteran plant pest, the whitefly Bemisia tabaci . Unlike Drosophila , where the paradigm for insect RNAi technology was established, whitefly has abundant somatic piwi-associated RNAs (piRNAs). Long regarded as germline restricted, piRNAs are common in the soma of many invertebrates. We sought to exploit this for a novel gene silencing approach. The main principle of piRNA biogenesis is the recruitment of target RNA fragments into the pathway. As such, we designed synthetic RNAs to possess complementarity to the loci we annotated. Following feeding of these exogenous piRNA triggers knockdown as effective as conventional siRNA-only approaches was observed. These results demonstrate a new approach for RNAi technology that could be applicable to dsRNA-recalcitrant pest species and could be fundamental to realizing insecticidal RNAi against pests. 
    more » « less
  3. null (Ed.)
    As an overarching immune mechanism, RNA interference (RNAi) displays pathogen specificity and memory via different pathways. The small interfering RNA (siRNA) pathway is the primary antiviral defense mechanism against RNA viruses of insects and plays a lesser role in defense against DNA viruses. Reflecting the pivotal role of the siRNA pathway in virus selection, different virus families have independently evolved unique strategies to counter this host response, including protein-mediated, decoy RNA–based, and microRNA-based strategies. In this review, we outline the interplay between insect viruses and the different pathways of the RNAi antiviral response; describe practical application of these interactions for improved expression systems and for pest and disease management; and highlight research avenues for advancement of the field. 
    more » « less
  4. Abstract Plant small RNAs are important regulatory elements that fine-tune gene expression and maintain genome integrity by silencing transposons. Reproductive organs of monocots produce abundant phased, small interfering RNAs (phasiRNAs). The 21-nt reproductive phasiRNAs triggered by miR2118 are highly enriched in pre-meiotic anthers, and have been found in multiple eudicot species, in contrast with prior reports of monocot specificity. The 24-nt reproductive phasiRNAs are triggered by miR2275, and are highly enriched during meiosis in many angiosperms. Here, we report the widespread presence of the 21-nt reproductive phasiRNA pathway in eudicots including canonical and non-canonical microRNA (miRNA) triggers of this pathway. In eudicots, these 21-nt phasiRNAs are enriched in pre-meiotic stages, a spatiotemporal distribution consistent with that of monocots and suggesting a role in anther development. Although this pathway is apparently absent in well-studied eudicot families including the Brassicaceae, Solanaceae and Fabaceae, our work in eudicots supports an earlier singular finding in spruce, a gymnosperm, indicating that the pathway of 21-nt reproductive phasiRNAs emerged in seed plants and was lost in some lineages. 
    more » « less
  5. Abstract Noncoding RNAs <200 nucleotides (nt) in length are referred to as short noncoding RNAs (sncRNAs) and include microRNAs (miRNAs), piwi-interacting RNAs, small nucleolar RNAs, transfer RNAs, etc. One striking example of the regulatory capabilities of sncRNAs comes from a group of small yet potent RNAs called miRNAs. MiRNAs are ∼20-nt RNAs excised from longer pre-miRNA hairpins, and to date, thousands of miRNAs have been identified across an array of species with specific roles for miRNAs defined in virtually every cellular activity (e.g. growth, differentiation, apoptosis, and disease). Importantly, studies aimed at evaluating the transcriptomic changes of miRNAs have now revealed the existence of miRNA-like fragments derived from other types of sncRNAs and suggest similar regulatory capacities may be associated with these novel sncRNA fragments. Unfortunately, many biologically relevant sncRNA-excised fragments remain uncharacterized due to their routine exclusion during initial miRNA characterizations as “sncRNA degradation products” as well as nearly all sncRNA informatic analyses continuing to solely assess annotated miRNA expressions. To address this, several platforms aimed at identifying novel sncRNA fragments have recently been developed. That said, the principal analytical tools currently employed to characterize novel sncRNA fragments often require significant computational expertise hindering their widespread utilization. As such, the development of a user-friendly platform, requiring minimal programming experience yet capable of identifying and characterizing RNA fragments excised from any sncRNA from any species is highly desirable and potentially impactful. In light of this, we have developed FragmentFinder—an intuitive, Windows-executable resource designed to require absolutely no computational background and capable of accurately characterizing all (annotated and unknown) sncRNA-derived RNAs within a raw small RNA sequencing file in real time. 
    more » « less