抗病毒基因HERC5新型环状RNA hsa＿circ＿0001426的特征分析及鉴定  

Characterization and Identification of a Novel Circular RNA Hsa＿circ＿0001426 Derived from Antiviral Gene HERC5

作　　者：姚文霞[1] 潘劲辉 梁雪 林海 梁敏[1] 刘兆宇[1] 周新科[1] YAO Wenxia;PAN Jinghui;LIANG Xue;LIN Hai;LIANG Min;LIU Zhaoyu;ZHOU Xinke(The Fifth Affiliated Hospital of Guangzhou Medical University,Guangzhou 510700,China)

机构地区：[1]广州医科大学附属第五医院,广东广州510700

出　　处：《现代医院》2019年第2期247-253,共7页Modern Hospitals

基　　金：国家自然科学基金项目(31500142; 81673206);高水平大学学术骨干培育计划(B185004047)

摘　　要：目的旨在研究来源于抗病毒基因HERC5的新型环状RNA hsa_circ_0001426的特征,并探索该环状RNA真核过表达载体的构建和鉴定。方法从反向剪切位点的验证、蛋白翻译的潜能、对RNase R处理的抵抗性、在细胞内的胞质胞核分布情况四个方面来研究hsa_circ_0001426的特征。此外,还利用环状RNA克隆构建试剂盒来构建hsa_circ_0001426的过表达载体,并验证其表达效果。结果 hsa_circ_0001426是由HERC5基因第18号外显子反向剪切至第12号外显子形成,其序列中包含一条可翻译318个氨基酸的开放阅读框,其抵抗Rnase R的处理,主要分布于细胞质。针对该环状RNA的过表达载体构建成功,其在转染HEK293T细胞后48 h的表达量最高。结论首次成功分析了抗病毒基因HERC5的新型环状RNA hsa_circ_0001426的特征,并利用试剂盒首次成功构建了hsa_circ_0001426的真核过表达载体,并可在细胞中高效表达;为深入研究hsa_circ_0001426的特征和功能奠定基础,并进一步扩充了HERC5基因的相关研究。Ojbective To characterize a novel circular RNA(circRNA)hsa_circ_0001426 derived from antiviral gene HERC5 and to explore the construction and identification of overexpression vector of hsa_circ_0001426.Methods The characteristics of hsa_circ_0001426 was studied from the following four aspects:verification of the backsplicing junction,analysis of the protein-coding ability,resistance to RNase R treatment,and the cellular distribution.In addition,we used commercially-available kits to construct overexpression vector of hsa_circ_0001426 and verify its expression effect.Results hsa_circ_0001426 was generated through backsplicing of exon 18 to exon 12 of HERC5 gene;there was a putative open reading frame(ORF)in hsa_circ_0001426 that encoded a 318 aa protein.Hsa_circ_0001426 was resistant to RNase R treatment and mainly localized in the cytoplasm.For the success of constructing overexpression vector of hsa_circ_0001426,the results of further transfection to HEK293T cells showed that hsa_circ_0001426 had maximum expression at 48h post transfection.Conclusion We successfully characterize the novel circRNA hsa_circ_0001426 derived from antiviral gene HERC5 for the first time;the eukaryotic overexpression vector of hsa_circ_0001426 was successfully constructed,which had high expression efficiency.This study lays a foundation for further study of the characteristics and functions of hsa_circ_0001426 and further expanded the study of HERC5 gene.

关 键 词：HERC5 环状RNA 蛋白翻译潜能 RNaseR抵抗性 胞质胞核分布 过表达 

分 类 号：R394-33[医药卫生—医学遗传学]