HSV-1即刻早期基因的5′UTR对报告基因mRNA翻译的影响  

Effect of the 5′untranslated region sequence of the HSV-1immediate early gene as the 5′UTR of the model protein mRNA on mRNA translation

作　　者：徐朗希 李丹丹 李恒 郑惠文[1] 彭沙沙 张莹[1] 于丽[1] 廖芸 刘龙丁[1] 李琦涵[1] XU Langxi;LI Dandan;LI Heng;ZHENG Huiwen;PENG Shasha;ZHANG Ying;YU Li;LIAO Yun;LIU Longding;LI Qihan(Institute of Medical Biology,Chinese Academy of Medical Sciences&Peking Union Medical College,Kunming 650118,Yunnan,China;Kunming Medical University,Kunming 650500,Yunnan,China)

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所,云南昆明650118 [2]昆明医科大学,云南昆明650500

出　　处：《云南大学学报（自然科学版）》2024年第5期980-992,共13页Journal of Yunnan University(Natural Sciences Edition)

基　　金：国家自然科学基金面上项目(82171817);云南省创新疫苗技术与产业转化研发平台(202002AA100009);云南省技术创新人才培养对象项目(202305AD160006).

摘　　要：mRNA疫苗的5′非翻译区(5’untranslated regions,5′UTR)对其表达起着重要作用,报告基因增强绿色荧光蛋白基因(enhanced green fluorescent protein,EGFP)用于检测5′UTR调控mRNA表达已经是一个较为成熟的实验方法.为研究单纯疱疹病毒1型(herpes simplex virus 1,HSV-1)即刻早期基因的5′UTR作为报告基因EGFP mRNA的5′UTR对其翻译的影响,本研究将4个即刻早期基因的5’UTR以及表达能力较强的人β珠蛋白基因(humanβGlobin,hBg)5′UTR插入报告基因EGFP 5’端,通过观察绿色荧光强度研究不同即刻早期基因5′UTR对mRNA翻译的影响.结果表明:经mRNA转染试剂和纳米脂质体颗粒分别递送进293T细胞和小鼠体内;RL2的5′UTR作为报告基因EGFP 5′UTR的表达水平与hBg 5′UTR相近,RS1、US1、US12表达水平明显弱于hBg 5′UTR.因此,RL2基因的5′UTR有潜力作为mRNA疫苗的5′UTR.The 5′untranslated region(5′UTR)of mRNA plays a crucial role in its expression.Using the enhanced green fluorescent protein gene(EGFP)as a reporter gene to detect the regulation of mRNA expression by 5′UTR is a well-established experimental method.This study aims to investigate the impact of the 5′UTR of herpes simplex virus 1(HSV-1)immediate-early genes on the translation of EGFP mRNA when used as its 5′UTR.To achieve this,the 5′UTRs of four immediate-early genes and the strongly expressive humanβ-globin gene(hBg)5′UTR were inserted at the 5′end of the reporter gene EGFP.By observing the intensity of green fluorescence,the effect of different immediate-early gene 5′UTRs on mRNA translation was studied.The results showed that,when delivered into 293T cells and mice using mRNA transfection reagents and nanoliposome particles,the expression level of EGFP with the RL25′UTR was similar to that of the hBg 5′UTR,while the expression levels of RS1,US1,and US12 were significantly weaker than that of the hBg 5′UTR.Therefore,the RL2 gene's 5′UTR has potential as a 5′UTR for mRNA vaccines.

关 键 词：单纯疱疹病毒1型 5′UTR mRNA 蛋白表达水平 

分 类 号：R186[医药卫生—流行病学]