Loading...
This record has been deleted. Since it is linked to a published record, it is available here for reference purposes only.
Click here to view the deleted recordThis document has been updated. This is not the latest published version. Click here to view the latest version of the record.
Living Modified Organism
(LMO)
The image below identifies the LMO through its unique identifier, trade name and a link to this page of the BCH. Click on it to download a larger image on your computer. For help on how to use it go to the LMO quick-links page.
GloFish® Electric Green® Tetra
EN
CGT2016 line
No
-
Organization:Yorktown Technologies ()Phone:Fax:Email:Website: http://www.glofish.com/,
The white tetra (Gymnocorymbus ternetzi) was modified to exhibit a permanent green coloration in natural light and fluorescent green colouration in blue or UV light. To achieve this color change from white to green, GloFish® Electric Green® expresses the green fluorescent protein gene from Zoanthus reef coral species.
The term “Recipient organism” refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas “Parental organisms” refers to those that were involved in cross breeding or cell fusion.
-
BCH-ORGA-SCBD-281063-2 Organism Gymnocorymbus ternetzi (Tetra, Black tetra, Black skirt tetra, Black widow, Petticoat tetra, Albino black skirt tetra, White tetra, White skirt tetra)Fish
Original species name is Tetragonopterus ternetzi Boulenger, 1895, the lectotype is in the Natural History Museum, London (BMNH : 1885.5.17.163). The individual fish used to produce CGT2016 were sourced from an ornamental aquarium fish producer (5-D Tropical) in 2007. Sperm from this line is deposited at the European Collection of Cell Cultures (ECACC), United Kingdom, as "Green tetra 1 transformation event" (accession no. 12061301).
EN
pZMLC-SV40X2-ZsGreen1
EN
- Microinjection
|
1.994 kb
|
|
0.716 kb
|
|
0.000 kb
|
|
0.000 kb
|
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
-
BCH-GENE-SCBD-114748-3 SV40 poly-adenylation signal | Macaca mulatta polyomavirus 1 (SV40, Simian vacuolating virus 40, simian virus 40, Rhesus macaque polyomavirus)Terminator
-
BCH-GENE-SCBD-281415-1 Myosin light chain promoter | Danio rerio (Zebrafish, Leopard danio, Zebra danio)Promoter
-
BCH-GENE-SCBD-281470-1 Green fluorescent protein gene 1 | Zoanthus sp. (green polyp, reef coral, mat anemone, button polyp)Protein coding sequence | Changes in physiology and/or production,Selectable marker genes and reporter genes
Information regarding the inserted DNA sequences
DNA constructs were built and microinjected into the white tetra embryos at a single insertion site.
Fluorescent proteins are expressed in the skeletal muscle of the modified tetra and are readily visible under normal daylight.
The transforming plasmid contains a cassette expressing the green fluorescent protein gene ZsGreen1 from the Zoanthus reef coral species. This cassette is under the control of a broadly strong, constitutive zebrafish (Danio rerio) myosin light chain promoter and two copies of the SV40 polyadenylation signals derived from Macaca mulatta polyomavirus 1. The myosin light chain promoter is muscle-specific, and the SV40 polyadenylation signals direct the 3′ end adenylation of the mRNA.
DNA constructs were built and microinjected into the white tetra embryos at a single insertion site.
Fluorescent proteins are expressed in the skeletal muscle of the modified tetra and are readily visible under normal daylight.
The transforming plasmid contains a cassette expressing the green fluorescent protein gene ZsGreen1 from the Zoanthus reef coral species. This cassette is under the control of a broadly strong, constitutive zebrafish (Danio rerio) myosin light chain promoter and two copies of the SV40 polyadenylation signals derived from Macaca mulatta polyomavirus 1. The myosin light chain promoter is muscle-specific, and the SV40 polyadenylation signals direct the 3′ end adenylation of the mRNA.
Note
- The SV40 polyadenylation signals in the vector pZMLC-SV40X2-ZsGreen1 are both 440 bp in total, with each signal ranging individually from 220 to 250 bp.
- Southern blot, quantitative PCR, and sequencing analysis confirmed that a single copy of the transgene expression cassette was integrated at a single locus, with no detectable plasmid backbone sequences present in the CGT2016 genome.
- Confirmation of a single transgene copy inserted at a single site with Mendelian segregation was also confirmed at the F3 generation.
EN
- Ornamental
EN
- Both the normal and long-fin white variants of the Black Tetra were used to produce CGT2016 line (white tetra was used due to the lack of dark striped pigmentation and would result in better fluorescence for ornamental purposes).
- The attached paper on the detection method (Debode et al., 2020) describes the detection of the green fluorescent protein (GFP) coding sequence from the Aequorea victoria jellyfish species. However, BLAST analysis of the primers "gloGreen-F" and "gloGreen-R" revealed a 100% sequence similarity to the ZsGreen1 fluorescent protein from Zoanthus reef coral species. Therefore, for the purposes of this record, the latter species is used as the reference for the GFP coding sequence.
- Two non-targeted phenotypic effects have been identified in CGT2016: reduced tolerance to low temperatures compared to wild-type siblings, and a more than 40% reduction in expected reproductive success in competition with wild-type siblings. However, there was no difference in juvenile survival.
- The GloFish® brand of the first fluorescent fish was created by Alan Blake and Richard Crockett in 2001 for the aquarium trade. GloFish® debuted in the United States in late 2003 as the first modified animal to become commercially available as a pet. The market then also reached Canada in late 2017.
- Patent: US20130333060A1 Green Transgenic Fluorescent Ornamental Fish [PDF] ( patent, ornamental fish, transgenic, green, fluorescent ) [ English ]
- Patent: US8975467 Green Transgenic Fluorescent Ornamental Fish [PDF] ( patent, ornamental fish, transgenic, green, florescent ) [ English ]
- Patent: US20090025645A1 Recombinant Constructs and Transgenic Ornamental Fish Therefrom [PDF] ( patent, recombinant constructs, fluorescent, ornamental fish ) [ English ]
- Canadian Science Advisory Secretariat: Environmental Risk Assessment of the GloFish Electric Green Tetra and the GloFish Long-Fin Electric Green Tetra [PDF] ( glofish, green, long-fin, fin, risk assessment ) [ English ]
- Canadian Science Advisory Secretariat: Environmental Protection Act Indirect Human Health Assessment [PDF] ( human health, assessment, tetra, glofish, electric green ) [ English ]
- Fish Physiology and Biochemistry (2019): Fluorescent protein transgenesis has varied effects on behaviour and cold tolerance in a tropical fish (Gymnocorymbus ternetzi): implications for risk assessment [DOI] [ English ]
- GloFish® official website [URL] [ English ]
| Record type | Field | Record(s) | |
|---|---|---|---|
| Country's Decision or any other Communication | LMO identification | 1 | |
