PUBLICATION
Gene delivery available in molluscan cells by strong promoter discovered from bivalve-infectious virus
- Authors
- Yoon, J., Gu, W.B., Konuma, M., Kobayashi, M., Yokoi, H., Osada, M., Nagasawa, K.
- ID
- ZDB-PUB-221103-8
- Date
- 2022
- Source
- Proceedings of the National Academy of Sciences of the United States of America 119: e2209910119 (Journal)
- Registered Authors
- Yokoi, Hayato
- Keywords
- CMV promoter, OsHV-1, electroporation, marine invertebrates, primary culture
- MeSH Terms
-
- Animals
- Bivalvia*
- HEK293 Cells
- Humans
- Promoter Regions, Genetic/genetics
- Zebrafish*
- PubMed
- 36322729 Full text @ Proc. Natl. Acad. Sci. USA
Citation
Yoon, J., Gu, W.B., Konuma, M., Kobayashi, M., Yokoi, H., Osada, M., Nagasawa, K. (2022) Gene delivery available in molluscan cells by strong promoter discovered from bivalve-infectious virus. Proceedings of the National Academy of Sciences of the United States of America. 119:e2209910119.
Abstract
Understanding gene functions in marine invertebrates has been limited, largely due to the lack of suitable assay systems. Such a system requires investigative methods that are reproducible and can be quantitatively evaluated, such as a cell line, and a strong promoter that can drive high expression of a transgene. In this study, we established primary cell culture from a marine bivalve mollusc, Mizuhopecten yessoensis. Using scallop primary cells, we optimized electroporation conditions for transfection and carried out a luciferase-based promoter activity assay to identify strong promoter sequences that can drive expression of a gene of interest. We evaluated potential promoter sequences from genes of endogenous and exogenous origin and discovered a strong viral promoter derived from a bivalve-infectious virus, ostreid herpesvirus-1 (OsHV-1). This promoter, we termed OsHV-1 promoter, showed 24.7-fold and 16.1-fold higher activity than the cytomegalovirus immediate early (CMV IE) promoter and the endogenous EF1α promoter, the two most commonly used promoters in bivalves so far. Our GFP assays showed that the OsHV-1 promoter is active not only in scallop cells but also in HEK293 cells and zebrafish embryos. The OsHV-1 promoter practically enables functional analysis of marine molluscan genes, which can contribute to unveiling gene-regulatory networks underlying astonishing regeneration, adaptation, reproduction, and aging in marine invertebrates.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping