title: Development of non-defective recombinant densovirus vectors for microRNA delivery in the invasive vector mosquito, Aedes albopictus.
abstract: We previously reported that mosquito densoviruses (MDVs) are potential vectors for delivering foreign nucleic acids into mosquito cells. However, considering existing expression strategies, recombinant viruses would inevitably become replication-defective viruses and lose their ability for secondary transmission. The packaging limitations of the virion represent a barrier for the development of MDVs for viral paratransgenesis or as high-efficiency bioinsecticides. Herein, we report the development of a non-defective recombinant Aedes aegypti densovirus (AaeDV) miRNA expression system, mediated by an artificial intron, using an intronic miRNA expression strategy. We demonstrated that this recombinant vector could be used to overexpress endogenous miRNAs or to decrease endogenous miRNAs by generating antisense sponges to explore the biological functions of miRNAs. In addition, the vector could express antisense-miRNAs to induce efficient gene silencing in vivo and in vitro. The recombinant virus effectively self-replicated and retained its secondary transmission ability, similar to the wild-type virus. The recombinant virus was also genetically stable. This study demonstrated the first construction of a non-defective recombinant MDV miRNA expression system, which represents a tool for the functional analysis of mosquito genes and lays the foundation for the application of viral paratransgenesis for dengue virus control.
authors.affiliation: Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, Guangdong, 510515, China.
authors.forename: Peiwen
authors.initials: P
authors.lastname: Liu
authors.affiliation: Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, Guangdong, 510515, China.
authors.forename: Xiaocong
authors.initials: X
authors.lastname: Li
authors.affiliation: Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, Guangdong, 510515, China.
authors.forename: Jinbao
authors.initials: J
authors.lastname: Gu
authors.affiliation: Reproductive Medical Center of Guangdong Women and Children Hospital, Guangzhou, Guangdong, 511442, China.
authors.forename: Yunqiao
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, Guangdong, 510515, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, Guangdong, 510515, China.
authors.forename: Puthiyakunnon
authors.initials: P
authors.lastname: Santhosh
authors.affiliation: Guangdong Provincial Key Laboratory of Tropical Disease Research, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, Guangdong, 510515, China.
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Chen
mesh_terms: D000330:Aedes / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017990:Densovirus / Q000235:genetics*; D015870:Gene Expression; D023061:Gene Order; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D042822:Genomic Instability; D007438:Introns; D035683:MicroRNAs / Q000737:chemistry / Q000235:genetics*; D034622:RNA Interference; D012326:RNA Splicing; D014161:Transduction, Genetic; D014779:Virus Replication
chemical_list: D035683:MicroRNAs