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首页> 外文期刊>Journal of Helminthology >Marker genes for activation of the RNA interference (RNAi) pathway in the free-living nematode Caenorhabditis elegans and RNAi development in the ovine nematode Teladorsagia circumcincta
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Marker genes for activation of the RNA interference (RNAi) pathway in the free-living nematode Caenorhabditis elegans and RNAi development in the ovine nematode Teladorsagia circumcincta

机译：自由活动线虫秀丽隐杆线虫RNA干扰（RNAi）途径的激活和绵羊线虫Teladorsagia circumcincta中RNAi发育的标记基因

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The nematode Teladorsagia circumcincta is a major cause of parasitic gastroenteritis in sheep in temperate regions. The development of resistance to the major anthelmintic classes used for its control is a threat to small ruminant farming sustainability. Vaccination is a potential alternative control method for this nematode. Gene datasets can be exploited to identify potential vaccine candidates and these validated further by methods such as RNA interference (RNAi) prior to vaccine trials. Previous reports indicate that RNAi in parasitic nematodes is inconsistent and, to date, there are no internal controls that indicate activation of the RNAi pathway in response to double-stranded RNA (dsRNA). The present aims were to determine whether or not the transcription levels of potential marker genes in the RNAi pathway could indicate activation of the pathway in Caenorhabditis elegans and to develop an RNAi platform in T. circumcincta. In C. elegans, transcript levels of three candidate marker genes, Ce-dcr-1 (Dicer), Ce-ego-1 (Enhancer of Glp-One family member) and Ce-rsd-3 (RNAi Spreading Defective), were analysed and results indicated that activation of the pathway had no effect on transcript levels of these genes. In T. circumcincta, two vaccine candidate genes from the Activation-associated Secreted Protein (ASP) family were targets for knockdown. RNAi experiments showed successful silencing of both targets, although inconsistencies in efficacy were observed. After testing a number of parameters that might affect variability, it was found that the length of the storage period of the larvae plays an important role in the consistency of the RNAi results.
机译：线虫Teladorsagia circumcincta是温带地区绵羊寄生性肠胃炎的主要原因。对用于控制其的主要驱虫类的抗药性的发展对反刍动物的农业可持续性构成了威胁。疫苗接种是该线虫的潜在替代控制方法。可以利用基因数据集来识别潜在的候选疫苗，并在疫苗试验之前通过诸如RNA干扰（RNAi）等方法进一步验证这些候选基因。先前的报道表明寄生线虫中的RNAi不一致，并且迄今为止，还没有内部对照表明响应双链RNA（dsRNA）的RNAi途径的激活。本发明的目的是确定RNAi途径中潜在标志物基因的转录水平是否可以表明秀丽隐杆线虫中该途径的活化，并开发出在包皮圆线虫中的RNAi平台。在秀丽隐杆线虫中，分析了三个候选标记基因Ce-dcr-1（切酶），Ce-ego-1（Glp-One家族成员的增强子）和Ce-rsd-3（RNAi传播缺陷）的转录水平。结果表明该途径的激活对这些基因的转录水平没有影响。在Circumcincta中，来自激活相关分泌蛋白（ASP）家族的两个候选疫苗基因是敲除的目标。 RNAi实验显示两个靶标均成功沉默，尽管观察到功效不一致。在测试了许多可能影响变异性的参数后，发现幼虫的储存期长度在RNAi结果的一致性中起着重要作用。

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