PMID- 30894668
OWN - NLM
STAT- MEDLINE
DCOM- 20201015
LR  - 20210109
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 9
IP  - 1
DP  - 2019 Mar 20
TI  - Comprehensive Experimental System for a Promising Model Organism Candidate for 
      Marine Teleosts.
PG  - 4948
LID - 10.1038/s41598-019-41468-8 [doi]
LID - 4948
AB  - A comprehensive experimental system for Japanese anchovy, a promising candidate 
      model organism for marine teleosts, was established. Through the design of a 
      rearing/spawning facility that controls the photoperiod and water temperature, 
      one-cell eggs were continuously obtained shortly after spawning throughout the 
      rearing period. The stages of eggs are indispensable for microinjection 
      experiments, and we developed an efficient and robust microinjection system for 
      the Japanese anchovy. Embryos injected with GFP mRNA showed strong whole-body GFP 
      fluorescence and the survival rates of injected- and non-injected embryos were 
      not significantly different, 87.5% (28 in 32 embryos) and 90.0% (45 in 50 
      embryos), respectively. We verified that the Tol2 transposon system, which 
      mediates gene transfer in vertebrates, worked efficiently in the Japanese anchovy 
      using the transient transgenesis protocol, with GFP or DsRed as the reporter 
      gene. Finally, we confirmed that genome-editing technologies, namely 
      Transcription Activator-Like Effector Nucleases (TALEN) and Clustered Regulatory 
      Interspaced Short Palindromic Repeats (CRISPR)/Cas9, were applicable to the 
      Japanese anchovy. In practice, specific gene-disrupted fishes were generated in 
      the F(1) generation. These results demonstrated the establishment of a basic, yet 
      comprehensive, experimental system, which could be employed to undertake 
      experiments using the Japanese anchovy as a model organism for marine teleost 
      fish.
FAU - Sakaguchi, Keishi
AU  - Sakaguchi K
AD  - Fisheries Research Institute of Karatsu, Department of Joint Research, Faculty of 
      Agriculture, Kyushu University, Saga, 847-0132, Japan. 
      keishi_s@agr.kyushu-u.ac.jp.
FAU - Yoneda, Michio
AU  - Yoneda M
AD  - National Research Institute of Fisheries and Environment of Inland Sea, 
      Hakatajima Station, Imabari, 794-2305, Japan.
FAU - Sakai, Noriyoshi
AU  - Sakai N
AD  - Genetic Strains Research Center, National Institute of Genetics, Mishima, 
      411-8540, Japan.
AD  - Department of Genetics, School of Life Sciences, SOKENDAI (The Graduate 
      University for Advanced Studies), Mishima, 411-8540, Japan.
FAU - Nakashima, Kanako
AU  - Nakashima K
AD  - Laboratory of Marine Biology, Department of Bioresource Sciences, Faculty of 
      Agriculture, Kyushu University, Fukuoka, 819-0385, Japan.
FAU - Kitano, Hajime
AU  - Kitano H
AD  - Fisheries Research Institute of Karatsu, Department of Joint Research, Faculty of 
      Agriculture, Kyushu University, Saga, 847-0132, Japan.
FAU - Matsuyama, Michiya
AU  - Matsuyama M
AD  - Laboratory of Marine Biology, Department of Bioresource Sciences, Faculty of 
      Agriculture, Kyushu University, Fukuoka, 819-0385, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190320
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (DNA Transposable Elements)
RN  - EC 3.1.- (Transcription Activator-Like Effector Nucleases)
SB  - IM
MH  - Animals
MH  - Animals, Genetically Modified
MH  - CRISPR-Cas Systems/genetics
MH  - DNA Transposable Elements/genetics
MH  - Embryo, Nonmammalian
MH  - Fishes/*physiology
MH  - Gene Editing/methods
MH  - Microinjections/methods
MH  - *Models, Animal
MH  - Seawater
MH  - Transcription Activator-Like Effector Nucleases/genetics
PMC - PMC6426966
COIS- The authors declare no competing interests.
EDAT- 2019/03/22 06:00
MHDA- 2020/10/21 06:00
PMCR- 2019/03/20
CRDT- 2019/03/22 06:00
PHST- 2019/01/08 00:00 [received]
PHST- 2019/03/08 00:00 [accepted]
PHST- 2019/03/22 06:00 [entrez]
PHST- 2019/03/22 06:00 [pubmed]
PHST- 2020/10/21 06:00 [medline]
PHST- 2019/03/20 00:00 [pmc-release]
AID - 10.1038/s41598-019-41468-8 [pii]
AID - 41468 [pii]
AID - 10.1038/s41598-019-41468-8 [doi]
PST - epublish
SO  - Sci Rep. 2019 Mar 20;9(1):4948. doi: 10.1038/s41598-019-41468-8.