Purple transgenic fluorescent ornamental fish
Inventors
Blake, Alan • Crockett, Richard • Nasevicius, Aidas
Assignees
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Publication Number
US-8232450-B1
Publication Date
2012-07-31
Expiration Date
Abstract
The present invention relates to the method and use of fluorescent proteins in making purple transgenic fluorescent fish. Also disclosed are methods of establishing a population of such transgenic fish and methods of providing them to the ornamental fish industry for the purpose of marketing. Thus, new varieties of ornamental fish of different fluorescence colors from a novel source are developed.
Core Innovation
The invention relates to purple fluorescent ornamental transgenic fish, specifically transgenic zebrafish (Danio rerio) that exhibit a fluorescence from a chromosomally integrated expression cassette encoding an FP635 fluorescent protein. The transgenic zebrafish comprise the Purple zebrafish 1 transformation event, where sperm comprising the Purple zebrafish 1 transformation event is deposited as ECACC accession no. 11012801.
The disclosed transgenic zebrafish are generated and characterized around a chromosomally integrated FP635 expression cassette associated with Purple zebrafish 1, including identification and breeding of transgenic lines to produce progeny. The document also describes sperm-based fertilization, including use of frozen sperm, and breeding to provide transgenic zebrafish comprising the Purple zebrafish 1 transformation event.
An ornamental base genotype is described via Golden zebrafish, involving a slc24a5 mutation associated with reduced melanosomes and lighter pigmentation, characterized as advantageous for producing an ornamental purple fluorescent fish. The document further addresses constructing the expression cassette with promoters, regulatory motifs, and FP635 fluorescent protein coding sequences, and distributing fluorescent fish to the ornamental industry, with Purple zebrafish 1 used as a stably expressing line.
Claims Coverage
The independent claims define two inventive features: a fluorescent transgenic zebrafish with a chromosomally integrated FP635 expression cassette associated with the Purple zebrafish 1 transformation event deposited under ECACC accession no. 11012801, and a method of producing such a transgenic fluorescent zebrafish by breeding. The claim set centers on the Purple zebrafish 1 event and a chromosomally integrated FP635 expression cassette, with a production claim that obtains an event-bearing fluorescent zebrafish and breeds it with a second zebrafish to obtain progeny comprising the same transformation event.
Transgenic zebrafish with chromosomally integrated FP635 from Purple zebrafish 1 event
A transgenic fluorescent zebrafish that exhibits a fluorescence and comprises a chromosomally integrated expression cassette encoding an FP635 fluorescent protein, wherein the zebrafish comprises the Purple zebrafish 1 transformation event, sperm comprising the Purple zebrafish 1 transformation event having been deposited as ECACC accession no. 11012801.
Producing transgenic zebrafish by breeding Purple zebrafish 1 event carriers
A method of producing a transgenic fluorescent zebrafish that exhibits a fluorescence, comprising obtaining a zebrafish that exhibits a fluorescence and comprises a chromosomally integrated expression cassette encoding an FP635 fluorescent protein, wherein the zebrafish comprises the Purple zebrafish 1 transformation event, sperm comprising the Purple zebrafish 1 transformation event having been deposited as ECACC accession no. 11012801; and breeding the obtained zebrafish with a second zebrafish to provide a transgenic zebrafish comprising the Purple zebrafish 1 transformation event.
Overall, the claim coverage is anchored on the Purple zebrafish 1 transformation event deposited as ECACC accession no. 11012801 and a chromosomally integrated expression cassette encoding FP635, with a breeding claim that obtains an event-bearing fluorescent zebrafish and breeds it with a second zebrafish to obtain progeny comprising the same Purple zebrafish 1 event.
Stated Advantages
Golden zebrafish are described as providing an advantageous ornamental base due to slc24a5 mutation effects on melanosomes and pigmentation.
Documented Applications
Cell lineage/migration tracing using Purple zebrafish 1 as a stably expressing fluorescent line.
Mosaic labeling using Purple zebrafish 1.
Fish cancer models using Purple zebrafish 1.
Market and research uses associated with fluorescent fish distribution to the ornamental fish industry.
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