云南大学学报(自然科学版)2024,Vol.46Issue(5) :980-992.DOI:10.7540/j.ynu.20240023

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

徐朗希 李丹丹 李恒 郑惠文 彭沙沙 张莹 于丽 廖芸 刘龙丁 李琦涵
云南大学学报(自然科学版)2024,Vol.46Issue(5) :980-992.DOI:10.7540/j.ynu.20240023

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

徐朗希 1李丹丹 1李恒 1郑惠文 1彭沙沙 2张莹 1于丽 1廖芸 1刘龙丁 1李琦涵1
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作者信息

  • 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. 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;昆明医科大学,云南昆明 650500
  • 折叠

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EGFP5'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hBg5'UTR.因此,RL2 基因的5'UTR有潜力作为mRNA疫苗的5'UTR.

Abstract

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 human β-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 RL2 5'UTR was similar to that of the hBg 5'UTR,while the expression levels of RS1,US1,and US12were significantly weaker than that of thehBg5'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

Key words

HSV-1/5'UTR/mRNA/protein expression levels

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基金项目

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)

出版年

2024
云南大学学报(自然科学版)
云南大学

云南大学学报(自然科学版)

CSTPCDCSCD北大核心
影响因子:0.663
ISSN:0258-7971
参考文献量32
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