腐食酪螨microRNA及其对过敏原基因表达调控作用研究

腐食酪螨既是一种重要的储藏物害螨，也是室内过敏原的重要来源螨种之一。miRNAs已报道于30余种节肢动物体内，具有调控生长发育、参与宿主与病原体的交互作用、参与宿主免疫反应等功能。经检索miRBase 21.0，尚未收录房舍和储藏物螨类miRNAs。本研究计划收集腐食酪螨卵、幼螨、若螨、成螨，分别建立miRNAs文库并测序，获得其miRNA表达谱；分别建立转录组文库并测序，获得其基因表达谱，包括所有过敏原基因；通过生物信息学软件预测miRNAs与过敏原基因的相互关系后，用螨细胞培养、基因沉默miRNAs、qRT-PCR等手段验证miRNAs对过敏原基因靶向调控作用。研究有望全面解析腐食酪螨miRNAs表达谱、过敏原组成，深化人们对miRNAs功能和作用的认识，为针对miRNAs设计抑制剂防制螨类以及制备重组过敏原诊治过敏性疾病等奠定基础。

The mold mite Tyrophagus putrescentiae (Astigmata: Acaridae) resides in a wide variety of stored products, causing economic losses, but is also found in human living environments, where it is associated with asthma, perennial rhinitis, atopic dermatitis, eczema, and other allergic disorders. The allergenicity of mites like T. putrescentiae can be affected by the expression of genes encoding their allergens. Mounting evidence demonstrates the critical roles of small non-coding microRNAs (miRNAs) in regulation of gene expression across various species. To date, however, no miRNAs have been identified from house dust mites and storage mites. We hypothesize that miRNAs are critical to the expression of allergen-encoding genes from T. putrescentiae. Here, we propose to collect the eggs, larva, nymph, and adults of T. putrescentiae for next-generation sequencing and bioinformatics analyses that will enable development of transcriptome libraries to resolve gene expression profiles. miRNA libraries will also be constructed to resolve miRNAs expression profiles. We will predict the associations between these microRNAs and their target genes, particularly encoding allergens, using bioinformatics tools. To clarify the regulatory roles of these miRNAs on allergen expression, molecular tools including culture of mites, silencing of miRNAs, and qRT-PCR to measure gene expression will be applied. The successful completion of the proposed studies will identify the allergen composition and miRNAs of T. putrescentiae for the first time. Such findings will be helpful for understanding how miRNAs regulate expression of allergen-encoding genes. Further, this knowledge can provide a basis for the design of targeting molecules that may alter allergen expression and for the production of recombinant allergens to diagnose and treat allergic diseases.