title: A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni.
abstract: Schistosomiasis is a major neglected tropical disease caused by trematodes from the genus Schistosoma. Because schistosomes exhibit a complex life cycle and numerous mechanisms for regulating gene expression, it is believed that spliced leader (SL) trans-splicing could play an important role in the biology of these parasites. The purpose of this study was to investigate the function of trans-splicing in Schistosoma mansoni through analysis of genes that may be regulated by this mechanism and via silencing SL-containing transcripts through RNA interference. Here, we report our analysis of SL transcript-enriched cDNA libraries from different S. mansoni life stages. Our results show that the trans-splicing mechanism is apparently not associated with specific genes, subcellular localisations or life stages. In cross-species comparisons, even though the sets of genes that are subject to SL trans-splicing regulation appear to differ between organisms, several commonly shared orthologues were observed. Knockdown of trans-spliced transcripts in sporocysts resulted in a systemic reduction of the expression levels of all tested trans-spliced transcripts; however, the only phenotypic effect observed was diminished larval size. Further studies involving the findings from this work will provide new insights into the role of trans-splicing in the biology of S. mansoni and other organisms. All Expressed Sequence Tags generated in this study were submitted to dbEST as five different libraries. The accessions for each library and for the individual sequences are as follows: (i) adult worms of mixed sexes (LIBEST_027999: JZ139310 - JZ139779), (ii) female adult worms (LIBEST_028000: JZ139780 - JZ140379), (iii) male adult worms (LIBEST_028001: JZ140380 - JZ141002), (iv) eggs (LIBEST_028002: JZ141003 - JZ141497) and (v) schistosomula (LIBEST_028003: JZ141498 - JZ141974).
authors.affiliation: Grupo de Genômica e Biologia Computacional, Centro de Pesquisas René Rachou, Fiocruz, Belo HorizonteMG, Brasil.
authors.forename: Marina de Moraes
authors.initials: Mde M
authors.lastname: Mourão
authors.forename: Mainá
authors.initials: M
authors.lastname: Bitar
authors.forename: Francisco Pereira
authors.initials: FP
authors.lastname: Lobo
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Peconick
authors.forename: Priscila
authors.initials: P
authors.lastname: Grynberg
authors.forename: Francisco
authors.initials: F
authors.lastname: Prosdocimi
authors.forename: Michael
authors.initials: M
authors.lastname: Waisberg
authors.forename: Gustavo Coutinho
authors.initials: GC
authors.lastname: Cerqueira
authors.forename: Andréa Mara
authors.initials: AM
authors.lastname: Macedo
authors.forename: Carlos Renato
authors.initials: CR
authors.lastname: Machado
authors.forename: Timothy
authors.initials: T
authors.lastname: Yoshino
authors.forename: Glória Regina
authors.initials: GR
authors.lastname: Franco
mesh_terms: D000818:Animals; D020224:Expressed Sequence Tags; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics; D055785:Gene Knockdown Techniques; D015723:Gene Library; D007814:Larva; D008018:Life Cycle Stages / Q000235:genetics; D008297:Male; D010641:Phenotype; D012322:RNA Precursors / Q000235:genetics / Q000302:isolation & purification*; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering / Q000378:metabolism; D020038:RNA, Spliced Leader / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D012550:Schistosoma mansoni / Q000235:genetics* / Q000254:growth & development; D020040:Trans-Splicing / Q000235:genetics / Q000502:physiology*
chemical_list: D012322:RNA Precursors; D012330:RNA, Double-Stranded; D034741:RNA, Small Interfering; D020038:RNA, Spliced Leader
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