Generation of viperin-knockout zebrafish by CRISPR/Cas9-mediated genome engineering and the effect of this mutation under VHSV infection

Shanaka, K.A.S.N.; Jung, Sumi; Madushani, K.P.; Wijerathna, H.M.S.M.; Tharuka, M.D. Neranjan; Kim, Myoung-Jin; Lee, Jehee

doi:10.1016/j.fsi.2022.10.040

Cited by 7 publications

(5 citation statements)

References 52 publications

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“…Three-month-old male zebrafish, including WT and the viperin -/- fish ( 11 ), were gradually brought to the experimental temperature of 15°C by reducing the temperature from 28°C (2°C reduced/day). The fish were acclimatized at 15°C for 1 week and evaluated for low-temperature adaptation, and those with abnormal phenotypes or behaviors were removed.…”

Section: Methodsmentioning

confidence: 99%

“…Virus culture and propagation were performed as previously described ( 11 ). Acclimatized fish were injected with two doses of WT VHSV (genotype 4a) (high amount = 5 × 10 6 TCID 50 /fish and low dose =1 × 10 6 TCID 50 /fish) intraperitoneally.…”

Section: Methodsmentioning

confidence: 99%

“…Although the importance of cholesterol in VHSV propagation has been established ( 10 ), the mechanism by which cholesterol modulation occurs in fish needs to be better defined. In a previously published paper, we described the successful generation of viperin -/- zebrafish, emphasizing the importance of Viperin in larval defense against viruses ( 11 ). In this study, we analyzed the effects of Viperin on macrophage and neutrophil migration patterns in viperin -/- fish and cellular metabolism.…”

Section: Introductionmentioning

confidence: 99%

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Viperin mutation is linked to immunity, immune cell dynamics, and metabolic alteration during VHSV infection in zebrafish

Shanaka,

Jung,

Madushani

et al. 2023

Front. Immunol.

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Viperin is a prominent antiviral protein found in animals. The primary function of Viperin is the production of 3’-deoxy-3’,4’-didehydro-cytidine triphosphate (ddhCTP), an inhibitory nucleotide involved in viral RNA synthesis. Studies in mammalian models have suggested that ddhCTP interferes with metabolic proteins. However, this hypothesis has yet to be tested in teleost. In this study, the role of Viperin in regulating metabolic alterations during viral hemorrhagic septicemia virus (VHSV) infection was tested. When infected with VHSV, viperin-/- fish showed considerably higher mortality rates. VHSV copy number and the expression of the NP gene were significantly increased in viperin-/- fish. Metabolic gene analysis revealed significant differences in soda, hif1a, fasn, and acc expression, indicating their impact on metabolism. Cholesterol analysis in zebrafish larvae during VHSV infection showed significant upregulation of cholesterol production without Viperin. In vitro analysis of ZF4 cells suggested a considerable reduction in lipid production and a significant upregulation of reactive oxygen species (ROS) generation with the overexpression of viperin. Neutrophil and macrophage recruitment were significantly modulated in viperin-/- fish compared to the wild-type (WT) fish. Thus, we have demonstrated that Viperin plays a role in interfering with metabolic alterations during VHSV infection.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Viperin mutation is linked to immunity, immune cell dynamics, and metabolic alteration during VHSV infection in zebrafish

Shanaka,

Jung,

Madushani

et al. 2023

Front. Immunol.

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“…The targets of studies include lysosomes [ 170 , 171 ], endoplasmic reticulum [ 172 ], membranes [ 173 ], membrane channels [ 174 ], cytoskeleton [ 175 ], recombination processes [ 176 ], aneuploidy [ 177 ], expression regulation [ 178 ], and signaling [ 179 ]. There is research on the genetic basis of the organism’s response to oxidative stress [ 180 , 181 , 182 , 183 , 184 ], hypoxia [ 185 , 186 , 187 ], sodium ion uptake from the surrounding aquatic environment and ammonia excretion [ 188 , 189 , 190 ], inflammation processes [ 191 , 192 ], immunity [ 193 , 194 , 195 , 196 ] and disease resistance [ 197 , 198 , 199 , 200 , 201 ].…”

Section: Genome Editing In Salmonidae and Cyprinidae Aquaculture Fish...mentioning

confidence: 99%

“…The role of two zebrafish genes in resistance to viral diseases was studied using the CRISPR/Cas9 method: the viperin gene in relation to the viral hemorrhagic septicemia virus [ 197 ] and the rrm1 gene in relation to the nervous necrosis virus [ 338 ]. In both cases, mutants showed greater susceptibility to viruses than wild-type fish.…”

Section: Genome Editing In Salmonidae and Cyprinidae Aquaculture Fish...mentioning

confidence: 99%

In Search of a Target Gene for a Desirable Phenotype in Aquaculture: Genome Editing of Cyprinidae and Salmonidae Species

Orlova,

Ruzina,

Emelianova

et al. 2024

Genes

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Aquaculture supplies the world food market with a significant amount of valuable protein. Highly productive aquaculture fishes can be derived by utilizing genome-editing methods, and the main problem is to choose a target gene to obtain the desirable phenotype. This paper presents a review of the studies of genome editing for genes controlling body development, growth, pigmentation and sex determination in five key aquaculture Salmonidae and Cyprinidae species, such as rainbow trout (Onchorhynchus mykiss), Atlantic salmon (Salmo salar), common carp (Cyprinus carpio), goldfish (Carassius auratus), Gibel carp (Carassius gibelio) and the model fish zebrafish (Danio rerio). Among the genes studied, the most applicable for aquaculture are mstnba, pomc, and acvr2, the knockout of which leads to enhanced muscle growth; runx2b, mutants of which do not form bones in myoseptae; lepr, whose lack of function makes fish fast-growing; fads2, Δ6abc/5Mt, and Δ6bcMt, affecting the composition of fatty acids in fish meat; dnd mettl3, and wnt4a, mutants of which are sterile; and disease-susceptibility genes prmt7, gab3, gcJAM-A, and cxcr3.2. Schemes for obtaining common carp populations consisting of only large females are promising for use in aquaculture. The immobilized and uncolored zebrafish line is of interest for laboratory use.

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Zebrafish as a Model for Investigating Antiviral Innate Immunity

Wu,

Zheng,

Chang

2024

Methods in Molecular Biology

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Generation of viperin-knockout zebrafish by CRISPR/Cas9-mediated genome engineering and the effect of this mutation under VHSV infection

Cited by 7 publications

References 52 publications

Viperin mutation is linked to immunity, immune cell dynamics, and metabolic alteration during VHSV infection in zebrafish

Viperin mutation is linked to immunity, immune cell dynamics, and metabolic alteration during VHSV infection in zebrafish

In Search of a Target Gene for a Desirable Phenotype in Aquaculture: Genome Editing of Cyprinidae and Salmonidae Species

Zebrafish as a Model for Investigating Antiviral Innate Immunity

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