Characterization of white spot syndrome virus immediate-early gene promoters

Lin, Fanyu; Huang, He; Wei, Ke; Hou, Luhong; Li, Fang; Yang, Feng

doi:10.1099/vir.0.047274-0

Cited by 5 publications

(4 citation statements)

References 30 publications

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“…After we confirmed that it was possible to electroporate scallop cells with mRNA, we proceeded with electroporation with plasmid DNA to search for a strong promoter. Because many functional viral promoters are approximately of 500- to 700-bp length ( 19 – 21 , 56 ), we cloned roughly 1,000-bp 5′-upstream regions of OsHV-1 genes to include the full viral promoter sequences. In addition to six OsHV-1 genes, 5′-upstream regions of scallop endogenous my-actβ, my-ef1α , and my-ef1β and WSSV wsv-ie1 and wsv465 sequences were tested as potential promoters.…”

Section: Discussionmentioning

confidence: 99%

“…Pwsv-ie1 is known to be functional not only in shrimp and insect cells ( 24 ) but also in fish ( 58 ) and mammalian cells ( 59 ). Considering its high activity in different groups of animals, Pwsv-ie1 was included in the assay, together with Pwsv465, of which promoter activity was higher than Pwsv-ie1 in Sf9 cells ( 56 ).…”

Section: Discussionmentioning

confidence: 99%

“…The DNA sequences for the wsv-ie1 promoter and wsv465 promoter were generated by gene synthesis (Eurofins Genomics) based on the National Center for Biotechnology Information database ( 56 ), and −502 and −475 5′ upstream regions from the wsv-ie1 and wsv465 (herein referred to as Pwsv-ie1 and Pwsv465) start codons were cloned into PCR-linearized pNL1.1.…”

Section: Methodsmentioning

confidence: 99%

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Gene delivery available in molluscan cells by strong promoter discovered from bivalve-infectious virus

Yoon

Konuma

et al. 2022

Proc. Natl. Acad. Sci. U.S.A.

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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.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Gene delivery available in molluscan cells by strong promoter discovered from bivalve-infectious virus

Yoon

Konuma

et al. 2022

Proc. Natl. Acad. Sci. U.S.A.

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“…WSSV is a double-stranded circular DNA virus whose genes can be classified as immediate early (IE), delayed early, late, and very late. IE gene products are expressed prior to the replication cycle, encoding transcriptional factors under strong promoter regulation (Sánchez-Paz, 2010;Lin et al, 2013). Among these promoters is IE1, the promoter of the immediate early 1 gene (IE1, or WSV069), which is activated by several host crustacean transcription factors (Liu et al, 2007;Huang et al, 2010;Wang et al, 2011;Qiu et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Lentiviral-Transduced Ectopic Expression of Androgenic Hormone in a Crustacean Hematopoietic Primary Cell Culture

et al. 2021

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Novel monosex biotechnologies in crustacean aquaculture involve the induction of sex reversal through manipulations of the androgenic gland (AG), and its most prominent factor, the insulin-like androgenic gland (IAG) hormone, during early developmental stages. In the giant freshwater prawn Macrobrachium rosenbergii, all-female populations are desirable for aquaculture, since the females can be stocked at higher densities and exhibit more uniform growth than males. Recently, a novel biotechnology for all-female aquaculture was developed based on injection into post-larvae of suspended primary AG cells from mature M. rosenbergii males. However, this biotechnology depends on the availability of appropriate male donors and it requires delicate surgical procedures on the small endocrine AG to produce appropriate quantities of cells for the sex manipulation. We therefore established a new platform for the production of M. rosenbergii IAG (Mr-IAG) in hematopoietic cells. The method rests on the induction of ectopic Mr-IAG expression under the white spot syndrome virus (WSSV) immediate-early 1 (IE1) promoter via a novel lentiviral vector. Our results demonstrate that M. rosenbergii primary cells infected with the Mr-IAG lentiviruses are capable of transcription, translation, and secretion of Mr-IAG in culture. Our new platform, which produces easy-to-harvest cells in abundance, could replace the AG cells used in the first step of the above-mentioned biotechnology for all-female aquaculture and, importantly, pave the way for producing monosex populations in other edible crustacean aquaculture species. In addition, a lentiviral system for crustacean cells provides a useful tool for basic and applied research in crustacean species.

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The promoter of the white spot syndrome virus immediate-early gene WSSV108 is activated by the cellular KLF transcription factor

Liu

Kou

et al. 2015

Developmental & Comparative Immunology

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Characterization of white spot syndrome virus immediate-early gene promoters

Cited by 5 publications

References 30 publications

Gene delivery available in molluscan cells by strong promoter discovered from bivalve-infectious virus

Gene delivery available in molluscan cells by strong promoter discovered from bivalve-infectious virus

Lentiviral-Transduced Ectopic Expression of Androgenic Hormone in a Crustacean Hematopoietic Primary Cell Culture

The promoter of the white spot syndrome virus immediate-early gene WSSV108 is activated by the cellular KLF transcription factor

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