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A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni

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...

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Autores principales: Mourão, Marina de Moraes, Bitar, Mainá, Lobo, Francisco Pereira, Peconick, Ana Paula, Grynberg, Priscila, Prosdocimi, Francisco, Waisberg, Michael, Cerqueira, Gustavo Coutinho, Macedo, Andréa Mara, Machado, Carlos Renato, Yoshino, Timothy, Franco, Glória Regina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Instituto Oswaldo Cruz 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3970683/
https://www.ncbi.nlm.nih.gov/pubmed/24037192
http://dx.doi.org/10.1590/0074-0276108062013006
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author Mourão, Marina de Moraes
Bitar, Mainá
Lobo, Francisco Pereira
Peconick, Ana Paula
Grynberg, Priscila
Prosdocimi, Francisco
Waisberg, Michael
Cerqueira, Gustavo Coutinho
Macedo, Andréa Mara
Machado, Carlos Renato
Yoshino, Timothy
Franco, Glória Regina
author_facet Mourão, Marina de Moraes
Bitar, Mainá
Lobo, Francisco Pereira
Peconick, Ana Paula
Grynberg, Priscila
Prosdocimi, Francisco
Waisberg, Michael
Cerqueira, Gustavo Coutinho
Macedo, Andréa Mara
Machado, Carlos Renato
Yoshino, Timothy
Franco, Glória Regina
author_sort Mourão, Marina de Moraes
collection PubMed
description 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).
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spelling pubmed-39706832014-05-21 A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni Mourão, Marina de Moraes Bitar, Mainá Lobo, Francisco Pereira Peconick, Ana Paula Grynberg, Priscila Prosdocimi, Francisco Waisberg, Michael Cerqueira, Gustavo Coutinho Macedo, Andréa Mara Machado, Carlos Renato Yoshino, Timothy Franco, Glória Regina Mem Inst Oswaldo Cruz Articles 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). Instituto Oswaldo Cruz 2013-09 /pmc/articles/PMC3970683/ /pubmed/24037192 http://dx.doi.org/10.1590/0074-0276108062013006 Text en http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Articles
Mourão, Marina de Moraes
Bitar, Mainá
Lobo, Francisco Pereira
Peconick, Ana Paula
Grynberg, Priscila
Prosdocimi, Francisco
Waisberg, Michael
Cerqueira, Gustavo Coutinho
Macedo, Andréa Mara
Machado, Carlos Renato
Yoshino, Timothy
Franco, Glória Regina
A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni
title A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni
title_full A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni
title_fullStr A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni
title_full_unstemmed A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni
title_short A directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in Schistosoma mansoni
title_sort directed approach for the identification of transcripts harbouring the spliced leader sequence and the effect of trans-splicing knockdown in schistosoma mansoni
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3970683/
https://www.ncbi.nlm.nih.gov/pubmed/24037192
http://dx.doi.org/10.1590/0074-0276108062013006
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