First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxins

BACKGROUND: Loxoscelism is the envenomation caused by the bite of Loxosceles spp. spiders. It entails severe necrotizing skin lesions, sometimes accompanied by systemic reactions and even death. There are no diagnostic means and treatment is mostly palliative. The main toxin, found in several isofor...

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Autores principales:	Sapag,Amalia, Salinas-Luypaert,Catalina, Constenla-Muñoz,Carlos
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2014
Materias:	Antivenom Loxosceles laeta RNA aptamers SELEX SMD-Ll1 SMD-Ll2 Sphingomyelinase D
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100002
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spelling	oai:scielo:S0716-976020140001000022015-10-30First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxinsSapag,AmaliaSalinas-Luypaert,CatalinaConstenla-Muñoz,Carlos Antivenom Loxosceles laeta RNA aptamers SELEX SMD-Ll1 SMD-Ll2 Sphingomyelinase D BACKGROUND: Loxoscelism is the envenomation caused by the bite of Loxosceles spp. spiders. It entails severe necrotizing skin lesions, sometimes accompanied by systemic reactions and even death. There are no diagnostic means and treatment is mostly palliative. The main toxin, found in several isoforms in the venom, is sphingomyelinase D (SMD), a phospholipase that has been used to generate antibodies intended for medical applications. Nucleic acid aptamers are a promising alternative to antibodies. Aptamers may be isolated from a combinatorial mixture of oligonucleotides by iterative selection of those that bind to the target. In this work, two Loxosceles laeta SMD isoforms, Ll1 and Ll2, were produced in bacteria and used as targets with the aim of identifying RNA aptamers that inhibit sphingomyelinase activity. RESULTS: Six RNA aptamers capable of eliciting partial but statistically significant inhibitions of the sphingomyelinase activity of recombinant SMD-Ll1 and SMD-Ll2 were obtained: four aptamers exert ~17% inhibition of SMD-Ll1, while two aptamers result in ~25% inhibition of SMD-Ll2 and ~18% cross inhibition of SMD-Ll1. CONCLUSIONS: This work is the first attempt to obtain aptamers with therapeutic and diagnostic potential for loxoscelism and provides an initial platform to undertake the development of novel anti Loxoscelesvenom agents.info:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.47 20142014-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100002en10.1186/0717-6287-47-2
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Antivenom Loxosceles laeta RNA aptamers SELEX SMD-Ll1 SMD-Ll2 Sphingomyelinase D
spellingShingle	Antivenom Loxosceles laeta RNA aptamers SELEX SMD-Ll1 SMD-Ll2 Sphingomyelinase D Sapag,Amalia Salinas-Luypaert,Catalina Constenla-Muñoz,Carlos First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxins
description	BACKGROUND: Loxoscelism is the envenomation caused by the bite of Loxosceles spp. spiders. It entails severe necrotizing skin lesions, sometimes accompanied by systemic reactions and even death. There are no diagnostic means and treatment is mostly palliative. The main toxin, found in several isoforms in the venom, is sphingomyelinase D (SMD), a phospholipase that has been used to generate antibodies intended for medical applications. Nucleic acid aptamers are a promising alternative to antibodies. Aptamers may be isolated from a combinatorial mixture of oligonucleotides by iterative selection of those that bind to the target. In this work, two Loxosceles laeta SMD isoforms, Ll1 and Ll2, were produced in bacteria and used as targets with the aim of identifying RNA aptamers that inhibit sphingomyelinase activity. RESULTS: Six RNA aptamers capable of eliciting partial but statistically significant inhibitions of the sphingomyelinase activity of recombinant SMD-Ll1 and SMD-Ll2 were obtained: four aptamers exert ~17% inhibition of SMD-Ll1, while two aptamers result in ~25% inhibition of SMD-Ll2 and ~18% cross inhibition of SMD-Ll1. CONCLUSIONS: This work is the first attempt to obtain aptamers with therapeutic and diagnostic potential for loxoscelism and provides an initial platform to undertake the development of novel anti Loxoscelesvenom agents.
author	Sapag,Amalia Salinas-Luypaert,Catalina Constenla-Muñoz,Carlos
author_facet	Sapag,Amalia Salinas-Luypaert,Catalina Constenla-Muñoz,Carlos
author_sort	Sapag,Amalia
title	First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxins
title_short	First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxins
title_full	First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxins
title_fullStr	First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxins
title_full_unstemmed	First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxins
title_sort	first report of in vitro selection of rna aptamers targeted to recombinant loxosceles laeta spider toxins
publisher	Sociedad de Biología de Chile
publishDate	2014
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602014000100002
work_keys_str_mv	AT sapagamalia firstreportofinvitroselectionofrnaaptamerstargetedtorecombinantloxosceleslaetaspidertoxins AT salinasluypaertcatalina firstreportofinvitroselectionofrnaaptamerstargetedtorecombinantloxosceleslaetaspidertoxins AT constenlamunozcarlos firstreportofinvitroselectionofrnaaptamerstargetedtorecombinantloxosceleslaetaspidertoxins
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First report of in vitro selection of RNA aptamers targeted to recombinant Loxosceles laeta spider toxins

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