title: miRNA29 Promotes Viral Replication During Early Stage of PRRSV Infection In Vitro.
abstract: miRNAs are involved in various biological processes, such as host-virus interactions and antiviral immunity. In this study, we investigated the role of miR-29 on porcine reproductive and respiratory syndrome virus (PRRSV) replication and its target genes. At first, miR-29a/b-1/c expression was detected when porcine alveolar macrophages (PAMs) were infected with PRRSV at different infective doses by real time-quantitative polymerase chain reaction (RT-qPCR). The result showed that miR-29a/b-1 expression significantly increased after 6 h (p < 0.01), with the peak around 24 h, miR-29c expression in each period of PRRSV infection was very low. Then, pre-miR-29a/b-1 lentiviral vectors were constructed. Absolute RT-qPCR analysis showed that PAMs transfected with pre-miR-29a/b-1 lentiviral vectors significantly promoted PRRSV replication in PAM within 24 h (p < 0.01). The expression of the target genes (AKT3, TP53INP1, and RPS6KB1) of miR-29a significantly reduced (p < 0.01). Western blot analysis showed that AKT3 and TP53INP1 are reduced at miR-29a overexpression. To further validate the interaction between miR-29a and its target gene sites, the luciferase assay results demonstrated that miR-29a interacted with AKT3 3'UTR 1676 and 1261 sites, leading the inhibition of luciferase expression. Our findings support that miR-29a could promote PRRSV replication during early stage of virus infection in vitro and AKT3 could be the target gene of miR-29a.
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Mengjiao
authors.initials: M
authors.lastname: Zhou
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Chuanmin
authors.initials: C
authors.lastname: Li
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Chunyan
authors.initials: C
authors.lastname: Lu
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Zhang
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Yunzhi
authors.initials: Y
authors.lastname: Pan
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: College of Animal Science, Jilin University , Changchun, Jilin, People's Republic of China .
authors.forename: Boxing
authors.initials: B
authors.lastname: Sun
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D016676:Macrophages, Alveolar / Q000821:virology*; D035683:MicroRNAs / Q000378:metabolism*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000821:virology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000502:physiology*; D060888:Real-Time Polymerase Chain Reaction; D013552:Swine; D014779:Virus Replication
chemical_list: D020413:3' Untranslated Regions; D035683:MicroRNAs
keywords: AKT3; PAM; PRRSV; miRNA29; replication