title: Development of hybrid baculovirus vectors for artificial MicroRNA delivery and prolonged gene suppression.
abstract: MicroRNA (miRNA) plays essential roles in regulating gene expression, but miRNA delivery remains a hurdle, thus entailing a vector system for efficient transfer. Baculovirus emerges as a promising gene delivery vector but its inherent transient expression restricts its applications in some scenarios. Therefore, this study primarily aimed to develop baculovirus as a miRNA expression vector for prolonged gene suppression. We constructed recombinant baculoviruses carrying artificial egfp-targeting miRNA sequences within the miR155 backbone, which after expression by the cytomegalovirus promoter could knockdown the enhanced green fluorescent protein (EGFP) expression in a sequence- and dose-dependent manner. By swapping the mature miRNA sequences, the baculovirus miRNA shuttle effectively repressed the overexpression of endogenous TNF-α in arthritic synoviocytes without inducing apoptosis. To prolong the baculovirus-mediated expression, we further developed a hybrid baculovirus vector that exploited the Sleeping Beauty (SB) transposon for gene integration and sustained miRNA expression. The hybrid baculovirus vector that combined the miR155 scaffold and SB transposon effectively repressed the transgene expression for a prolonged period of time, hence diversifying the applications of baculovirus to indications necessitating prolonged gene regulation such as arthritis.
authors: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors: Chiu-Ling
authors: CL
authors: Chen
authors: Wen-Yi
authors: WY
authors: Luo
authors: Wen-Hsin
authors: WH
authors: Lo
authors: Kun-Ju
authors: KJ
authors: Lin
authors: Li-Yu
authors: LY
authors: Sung
authors: Yung-Shen
authors: YS
authors: Shih
authors: Yu-Han
authors: YH
authors: Chang
authors: Yu-Chen
authors: YC
authors: Hu
mesh_terms: D016367:Baculoviridae / Q000235:genetics*; D001688:Biological Products / Q000494:pharmacology*; D002460:Cell Line; D004251:DNA Transposable Elements; D004337:Drug Carriers; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005822:Genetic Vectors; D042822:Genomic Instability; D049452:Green Fluorescent Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000494:pharmacology*; D016254:Mutagenesis, Insertional; D011995:Recombination, Genetic; D014161:Transduction, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000235:genetics
chemical_list: D001688:Biological Products; D004251:DNA Transposable Elements; D004337:Drug Carriers; D035683:MicroRNAs; D014409:Tumor Necrosis Factor-alpha; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins