S

Glofish

GFP

Sources of GFP

GFP (Aequorea victoria, jellyfish),

GFP (Renilla reniformis, sea pansy),

dsRed (Discosoma, mushroom coral),

eqFP611 (Entacmaea quadricolor, sea anemone),

RTMS5 (Montipora efflorescens, stony coral),

dronpa (Pectiniidae, chalice coral),

KFP (Anemonia sulcata, Venus hair anemone),

eosFP (Lobophyllia hemprichii, open brain coral),

dendra (Dendronephthya, octocoral)

www.glofish.com

GloFish casts light on murkypolicing of transgenic animalsNature 27 November 2003

The New York Times Nov 22, 2003Gene-Altering RevolutionNears the Pet Store:Glow-in-the-Dark Fish

Marketed without regulatory environmental review. FDA is leading authority.

First Transgenic Animal in US market

GloFish

patented and trademarked brand of genetically modified (GM) fluorescent fish

variety of different GloFish are currently on the market

Zebrafish were the first GloFish available in pet stores

now sold in bright red, green, orange-yellow, blue, and purple fluorescent colors

Glofish retail for about $5 per fish. Normal zebrafish cost around one tenth of the price

Original Zebrafish

The original zebrafish (or zebra danio, Danio rerio) from which the GloFish was developed is a native of rivers in India and Bangladesh

Measures 3 cm long

Has gold and dark blue stripes

Over 200 million have been sold in the last 50 years in US

They have never established any wild populations because they are tropical, unable to survive in North America’s temperate climate

Researchers in Singapore added a fluorescence gene from a sea coral to zebra

danio eggs to produce glofish

In 1999, Dr. Zhiyuan Gong and his colleagues at the National University of Singapore worked with the gene encoding GFP or green fluorescent protein extracted from a jellyfish

They inserted the gene into zebrafish embryo and it integrated into the zebrafish genome

which caused the fish to be brightly fluorescent under both natural white light and ultraviolet light

Goal: develop a fish that could detect pollution by selectively fluoresce in the presence of environmental toxins

Developing a constantly fluorescing fish was the first step

Patent filed for their process

Soon other fluorescent zebra fish were developed using genes that produced different color: red from sea coral and orange-yellow from a variant of the jellyfish gene

Fluorescent medaka

Then a team of researchers at the National Taiwan University, headed by Professor Huai-Jen Tsai (蔡懷) succeeded in creating a medaka (rice fish) with a green fluorescent color as a model

A separate deal was made between Taikong, the largest aquarium produce in Taiwan and the Taiwanese researchers to market the green medaka in Taiwan under the name TK-1

In spring 2003, Taiwan became the first to authorize sales of a genetically modified organism as a pet. 100,000 were sold in less than a month at US$18.60 each

Yorktown Technologies

Deal between scientists from NUS (National University of Singapore) and businessmen Alan Blake and Richard Crockett from Yorktown Technologies, L.P., a company in Austin, Texas,

Yorktown obtained worldwide rights to market the fluorescent zebrafish, which Yorktown branded as "GloFish”

GloFish have been marketed in the United States since late 2003

there have been no reports of any ecological problems associated with their sale

Reproduction limited

speculation that the eggs are pressure treated to make them infertile

some GloFish are fertile and will reproduce in a captive environment

GloFish Fluorescent Fish License states "Intentional breeding and/or any sale, barter, or trade, of any offspring of GloFish fluorescent ornamental fish is strictly prohibited”

Reduced fertility

GloFish have reduced fertility compared to the species, possibly due to higher energy costs associated with fluorescence, instead of other bodily functions

Sale of Glofish restricted

Canada prohibits import or sale of the fish, due to what they report is a lack of sufficient information to make a decision with regard to safety

The import, sale and possession of these fish is not permitted within the European Union

Sale or possession of Glofish remains illegal in California due to a regulation that restricts all genetically modified fish, due to concern about a fast-growing biotech salmon

Glofish chemical detector

GloFish also have been used for experimental research.

The alterations in the zebrafish's genes has given the organism the ability to fluoresce as a bio-indicator.

This genetic ability has been used to detect pollution and other chemicals.

Chemicals that mimic natural estrogens have well-documented effects on the reproductive systems of invertebrates, typically acting as endocrine disruptors, and glofish flourescence is used to detect levels of estrogenic chemicals

Muscles such as the heart have a larger effect of estrogen than the liver.

History of Glofish

First - the red fluorescent zebrafish, trademarked as "Starfire Red”

Yorktown Technologies released a green fluorescent zebrafish and an orange-yellow fluorescent zebrafish in mid-2006

In 2011, blue and purple fluorescent zebrafish were released. These lines of fish are trademarked as "Electric Green", "Sunburst Orange", "Cosmic Blue", and "Galactic Purple", and incorporate genes from sea coral

In 2012, Yorktown Technologies introduced a new variety of "Electric Green" GloFish, derived from a different species of fish, the black tetra

Followed in 2013 by the “electric green” barb which is a variety of tiger barb and a “sunburst orange” tetra and a “moonrise pink” tetra, the first flourescent pink to be marketed (from YT)

Cosmic blue glofish

36085b

Starfire red zebra glofish

36055b red glo36055a red

Galactic purple zebra?

GloFish® Galactic Purple fluorescent fish bring color and excitement to any aquarium

A traditional white aquarium light will work for daytime viewing, but won’t give the fish its best color

An actinic light will give the fish an exciting and uniquely fluorescent look, particularly in low lighting environments - the color you look for in a glofish

If the room is completely dark, a black light will create the appearance that the fish are glowing in the dark

http://www.petsolutions.com/C/Live-Freshwater-Fish-Danios/I/Glofish-Cosmic-Blue-Zebra.aspx

Glofish: green tetra

GloFish® Electric Green Tetras are not injected or dyed. They breed and reproduce naturally with this fluorescent color.Electric Green Tetras look great under either regular daylight bulbs or under an actinic bulb.

Green Tetra GloFish® Electric Green Tetras are beautiful freshwater Tetra fish that

glow under black lights.

Unlike other GloFish® fluorescent fish, which are Zebra Danios, the Electric Green Tetra is a variation of the White Skirt Tetra. Electric Greens are created with selective breeding of the White Skirt Tetra, using a certain gene in the fish to produce the Electric Green color

One of the hardier Tetras, it prefers to school. They do best when there are more than 3 Electric Green Tetras in the school

While requiring some swimming space, Electric Greens will also appreciate cover along the perimeter of the aquarium

Generally peaceful, they can match up with just about any other type of fish that is not large enough to eat them

While the Electric Green Tetra might prefer softer, acidic water conditions, most can adapt to a wide range of water parameters and will do fine in a well-lit aquarium

Purposes of Transgenics

Industrial

Medical xenotransplantation nutritional supplements

and pharmaceuticals

Basic Research gene function gene expression

patterns

Agriculture size quality disease resistance

http://tasq.uq.edu.au/blasto.html

Blastocyst Injection

Blastocyst ES cells

TransgenicWildtype

http://www.nature.com/nbt/journal/v19/n6/images/nbt0601_500a_I1.jpg

Transgenic Fish

Tilapia

Salmon/trout

Catfish

Can grow up to 6 times faster than wildtype fish

Most have extra copies of growth hormone (GH) gene

Fish Transgenics

The transgene used to increase growth utilizes an antifreeze protein promoter connected to the GH cDNA

As water temperature drops the GH gene is turned on

The fish continue to grow when normally they would not

What happens if they escape?

Could become first transgenic animal approved for large-scale farming and human food

The New York Times

Age = 7.5 months

Transgenic = 1.2 kg

Unmodified = 200 g

Http://webhost.avin.net/afprotein/peidof.htm

Engineered with ocean pout antifreeze gene promoter + chinook salmon growth hormone gene