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微小RNA-339-5p通过靶向病毒基因区域抑制猪繁殖与呼吸综合征病毒的复制。

MiR-339-5p inhibits replication of porcine reproductive and respiratory syndrome virus by targeting viral gene regions.

作者信息

Ye Cuifang, Cao Xinyan, Sheng Jinliang, Sun Yanming, Li Guang, Fang Wenbin, Zhang Yanbing

机构信息

College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang, China.

Eighth Division, Animal Husbandry and Veterinary Workstation, Shihezi City, Xinjiang Production and Construction Corps, Shihezi, Xinjiang, China.

出版信息

Virus Genes. 2024 Apr;60(2):186-193. doi: 10.1007/s11262-024-02059-6. Epub 2024 Feb 18.

Abstract

Porcine reproductive and respiratory syndrome virus (PRRSV) is a variable virus, whose spread cannot be totally stopped by vaccination. PRRSV infection results in abortion and respiratory symptoms in pregnant pigs. One crucial component of the anti-viral infection strategy is microRNA (miRNA), a class of multifunctional small molecules. It is unknown whether miR-339-5p can specifically target the PRRSV gene and prevent the virus from replicating, despite the fact that miR-339-5p is markedly up-regulated during the PRRSV infection. In this pursuit, the present study revealed that the two PRRSV areas targeted by miR-339-5p were PRRSV nsp2-3378 to 3403 and PRRSV nsp2-3112 to 3133 using the miRanda program. Dual luciferase reporter assays showed that the miR-339-5p target region of the PRRSV gene sequence exhibited 100% homology and was highly conserved. Furthermore, the ability of miR-339-5p to target PRRSV gene areas was verified. It was found that the overexpression of miR-339-5p markedly reduced the PRRSV replication through PRRSV infection trials. The precursor sequence of ssc-miR-339-5p was amplified using the DNA of pig lung tissue as a template in order to create a fragment of 402 bp of porcine-derived miR-339-5p precursor sequence, which was then used to produce the eukaryotic expression plasmid of miR-339-5p. In conclusion, miR-339-5p can target the specific PRRSV gene areas and prevent PRRSV replication, offering fresh perspectives for the creation of medications that combat the PRRSV infection.

摘要

猪繁殖与呼吸综合征病毒(PRRSV)是一种变异病毒,疫苗接种无法完全阻止其传播。PRRSV感染会导致妊娠母猪流产和出现呼吸道症状。抗病毒感染策略的一个关键组成部分是微小RNA(miRNA),这是一类多功能小分子。尽管在PRRSV感染期间miR-339-5p明显上调,但尚不清楚它是否能特异性靶向PRRSV基因并阻止病毒复制。在这项研究中,使用miRanda程序发现miR-339-5p靶向的两个PRRSV区域是PRRSV nsp2-3378至3403以及PRRSV nsp2-3112至3133。双荧光素酶报告基因检测表明,PRRSV基因序列的miR-339-5p靶区域显示出100%的同源性且高度保守。此外,还验证了miR-339-5p靶向PRRSV基因区域的能力。通过PRRSV感染试验发现,miR-339-5p的过表达显著降低了PRRSV的复制。以猪肺组织DNA为模板扩增了ssc-miR-339-5p的前体序列,以产生一段402 bp的猪源miR-339-5p前体序列片段,然后用于制备miR-339-5p的真核表达质粒。总之,miR-339-5p可以靶向PRRSV的特定基因区域并阻止PRRSV复制,为开发对抗PRRSV感染的药物提供了新的视角。

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