title: Genome-wide analyses of proliferation-important genes of Iridovirus-tiger frog virus by RNAi.
abstract: Tiger frog virus (TFV), a species of genus Ranavirus in the family Iridoviridae, is a nuclear cytoplasmic large DNA virus that infects aquatic vertebrates such as tiger frog (Rana tigrina rugulosa) and Chinese soft-shelled turtle (Trionyx sinensis). Based on the available genome sequences of TFV, the well-developed RNA interference (RNAi) technique, and the reliable cell line for infection model, we decided to analyze the functional importance of all predicted genes. Firstly, a relative quantitative cytopathogenic effect (Q-CPE) assay was established to monitor the viral proliferation in fish cells. Then, genome-wide RNAi screens of 95 small interference (si) RNAs against TFV were performed to characterize the functional importance of nearly all (95%) predicted TFV genes by Q-CPE scaling system. We identified 32 (33.7%) genes as essential, 50 (52.6%) genes as semi-essential and 13 (13.7%) genes as nonessential for TFV proliferation. Quantitative RT-PCR and titer assays of selected genes were performed to verify the screen results. Furthermore, the screened essential genes were analyzed for their genome distribution and conservative comparison within Ranavirus. Such a systematic screen for viral functional genes by cell phenotypes should provide further insights into understanding of the information in antiviral targets, and in viral replication and pathogenesis of iridovirus.
authors.affiliation: State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
authors.forename: Jun-Feng
authors.initials: JF
authors.lastname: Xie
authors.affiliation: Department of Nephrology, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou 518080, China.
authors.forename: Yu-Xiong
authors.initials: YX
authors.lastname: Lai
authors.affiliation: State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
authors.forename: Li-Jie
authors.initials: LJ
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
authors.forename: Run-Qing
authors.initials: RQ
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
authors.forename: Shao-Wei
authors.initials: SW
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
authors.forename: Shao-Ping
authors.initials: SP
authors.lastname: Weng
authors.affiliation: State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Biocontrol/MOE Key Laboratory of Aquatic Product Safety, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China; School of Marine Sciences, Sun Yat-sen University, Guangzhou 510275, China. Electronic address: lsshjg@mail.sysu.edu.cn.
authors.forename: Jian-Guo
authors.initials: JG
authors.lastname: He
mesh_terms: D000818:Animals; D002460:Cell Line; D005399:Fishes; D023061:Gene Order; D020868:Gene Silencing; D020043:Genes, Essential; D005814:Genes, Viral; D017994:Iridovirus / Q000235:genetics / Q000502:physiology*; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D019562:Viral Load; D010948:Viral Plaque Assay; D014779:Virus Replication
keywords: Essential gene; Genome-wide RNAi; Iridovirus; Tiger frog virus; siRNA