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

机译:体外选择靶向重组人脊线虫毒素的RNA适体的首次报道

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摘要

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. 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. 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 Loxosceles venom agents.
机译:Loxoscelism是由Loxosceles spp的叮咬引起的毒液。蜘蛛。它导致严重的坏死性皮肤病变,有时伴有全身反应甚至死亡。没有诊断手段,治疗主要是姑息治疗。在毒液的几种同工型中发现的主要毒素是鞘磷脂酶D(SMD),这是一种磷脂酶,已被用于生成用于医学应用的抗体。核酸适体是抗体的有希望的替代物。可以通过迭代选择与靶标结合的寡核苷酸,从寡核苷酸的组合混合物中分离适体。在这项工作中,在细菌中产生了两种Lxosceles laeta SMD亚型Ll1和Ll2,并用作靶标,目的是鉴定抑制鞘磷脂酶活性的RNA适体。获得了六种能够引起重组SMD-Ll1和SMD-Ll2的鞘磷脂酶活性部分但统计学上显着抑制的RNA适体:四个适体对SMD-Ll1有约17%的抑制作用,而两个适体对SMD有约25%的抑制作用。 -Ll2和〜18%的SMD-Ll1交叉抑制。这项工作是获得具有治疗和诊断潜力的适配子的首次尝试,并提供了一个平台来进行新型抗蛇毒毒剂的开发。

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