PUBLICATION
Generation of viperin-knockout zebrafish by CRISPR/Cas9-mediated genome engineering and the effect of this mutation under VHSV infection
- Authors
- Shanaka, K.A.S.N., Jung, S., Madushani, K.P., Wijerathna, H.M.S.M., Neranjan Tharuka, M.D., Kim, M.J., Lee, J.
- ID
- ZDB-PUB-221030-4
- Date
- 2022
- Source
- Fish & shellfish immunology 131: 672-681 (Journal)
- Registered Authors
- Kim, Myoung-Jin
- Keywords
- CRISPR/Cas9, Knockout model, VHSV, Viperin, Zebrafish
- MeSH Terms
-
- Animals
- CRISPR-Cas Systems
- Fish Diseases*
- Hemorrhagic Septicemia, Viral*
- Interferons/genetics
- Mutation
- Novirhabdovirus*/physiology
- Viral Proteins/genetics
- Zebrafish/genetics
- Zebrafish/metabolism
- PubMed
- 36309322 Full text @ Fish Shellfish Immunol.
Citation
Shanaka, K.A.S.N., Jung, S., Madushani, K.P., Wijerathna, H.M.S.M., Neranjan Tharuka, M.D., Kim, M.J., Lee, J. (2022) Generation of viperin-knockout zebrafish by CRISPR/Cas9-mediated genome engineering and the effect of this mutation under VHSV infection. Fish & shellfish immunology. 131:672-681.
Abstract
Viperin is an important virus-induced protein in animals that negatively participates in RNA viral replication and transcription. The reactive machinery of viperin suggests that it produces the regulatory molecule ddhCTP, which may affect immune regulation. In this study, we investigated the expression pattern of viperin in the larval and adult stages of zebrafish by whole-mount in situ hybridization and reverse transcription-quantitative PCR (RT-qPCR). To elucidate the function of viperin, we generated a zebrafish knockout model using a CRISPR/Cas9 method and evaluated the mutation's effects under viral hemorrhagic septicemia virus (VHSV) infections. In zebrafish larvae, viperin was expressed in the brain region, eye, and pharynx, which was confirmed by cryosectioning. In adult zebrafish, blood cells showed the highest levels of viperin expression. In 5 dpf fish challenged with VHSV, the expression of the viral NP protein was significantly enhanced in viperin-/- fish compared to wild-type fish. In vitro VHSV propagation analysis indicated comparatively higher levels of virus propagation in viperin-/- fish. Mortality analysis confirmed higher mortality rates, and interferon gene expression analysis showed a strong upregulation of the interferon (ifn)φ1 and 3 gene in viperin-/- fish infected with VHSV. This study describes the successful generation of a viperin-knockout model and the role of viperin during VHSV infections.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping