PMID- 36322729
OWN - NLM
STAT- MEDLINE
DCOM- 20221104
LR  - 20230503
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 119
IP  - 45
DP  - 2022 Nov 8
TI  - Gene delivery available in molluscan cells by strong promoter discovered from 
      bivalve-infectious virus.
PG  - e2209910119
LID - 10.1073/pnas.2209910119 [doi]
LID - e2209910119
AB  - 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 EF1alpha 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.
FAU - Yoon, Jeongwoong
AU  - Yoon J
AUID- ORCID: 0000-0002-9031-8412
AD  - Laboratory of Aquaculture Biology, Graduate School of Agricultural Science, 
      Tohoku University, Sendai, Miyagi 980-8572, Japan.
FAU - Gu, Wen-Bin
AU  - Gu WB
AUID- ORCID: 0000-0002-8491-6539
AD  - Laboratory of Aquaculture Biology, Graduate School of Agricultural Science, 
      Tohoku University, Sendai, Miyagi 980-8572, Japan.
FAU - Konuma, Mizuki
AU  - Konuma M
AD  - Laboratory of Aquaculture Biology, Graduate School of Agricultural Science, 
      Tohoku University, Sendai, Miyagi 980-8572, Japan.
FAU - Kobayashi, Mutsuko
AU  - Kobayashi M
AD  - Laboratory of Aquaculture Biology, Graduate School of Agricultural Science, 
      Tohoku University, Sendai, Miyagi 980-8572, Japan.
FAU - Yokoi, Hayato
AU  - Yokoi H
AUID- ORCID: 0000-0003-1020-6160
AD  - Laboratory of Fish Genetics, Graduate School of Agricultural Science, Tohoku 
      University, Sendai, Miyagi 980-8572, Japan.
FAU - Osada, Makoto
AU  - Osada M
AUID- ORCID: 0000-0001-6247-5106
AD  - Laboratory of Aquaculture Biology, Graduate School of Agricultural Science, 
      Tohoku University, Sendai, Miyagi 980-8572, Japan.
FAU - Nagasawa, Kazue
AU  - Nagasawa K
AUID- ORCID: 0000-0003-2819-3302
AD  - Laboratory of Aquaculture Biology, Graduate School of Agricultural Science, 
      Tohoku University, Sendai, Miyagi 980-8572, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20221102
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - Mizuhopecten yessoensis
SB  - IM
MH  - Animals
MH  - Humans
MH  - *Zebrafish
MH  - HEK293 Cells
MH  - Promoter Regions, Genetic/genetics
MH  - *Bivalvia
PMC - PMC9661190
OTO - NOTNLM
OT  - CMV promoter
OT  - OsHV-1
OT  - electroporation
OT  - marine invertebrates
OT  - primary culture
COIS- The authors declare no competing interest.
EDAT- 2022/11/03 06:00
MHDA- 2022/11/05 06:00
PMCR- 2023/05/02
CRDT- 2022/11/02 15:03
PHST- 2022/11/02 15:03 [entrez]
PHST- 2022/11/03 06:00 [pubmed]
PHST- 2022/11/05 06:00 [medline]
PHST- 2023/05/02 00:00 [pmc-release]
AID - 202209910 [pii]
AID - 10.1073/pnas.2209910119 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2022 Nov 8;119(45):e2209910119. doi: 
      10.1073/pnas.2209910119. Epub 2022 Nov 2.