Red transgenic fluorescent ornamental fish
Inventors
Blake, Alan • Crockett, Richard • Nasevicius, Aidas
Assignees
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Publication Number
US-11202444-B2
Publication Date
2021-12-21
Expiration Date
Abstract
The present invention relates to transgenic red ornamental fish, as well as methods of making such fish by in vitro fertilization techniques. 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.
Core Innovation
The invention relates to transgenic red fluorescent ornamental fish and breeding methods. It specifically describes a “Red barb 1 transformation event” comprising chromosomally integrated fluorescent protein expression cassettes. The transgenic fish include barbs, including albino tiger barbs, Puntius tetrazona, that exhibit red fluorescence.
The transgenic fish and breeding materials are defined in terms of genetic and reproductive components carrying the “Red barb 1 transformation event.” Gametes include eggs, sperm, and embryos that carry chromosomally integrated expression cassettes encoding fluorescent protein. The sperm for the “Red barb 1 transformation event” is described as having been deposited as ECACC accession no. 16121901.
The disclosure further describes establishing transgenic populations and identifying individuals by red fluorescence. It also describes using frozen sperm and in vitro fertilization to generate transgenic progeny and hybrid progeny that carry the “Red barb 1 transformation event.” The resulting transgenic progeny are associated with ornamental markets and research tools, including cell lineage/migration tracing, genetic mosaic marking, and fish cancer models.
Claims Coverage
The claims cover one independent breeding method and dependent refinements. The inventive features center on breeding barbs to obtain a third transgenic barb hybrid carrying the “Red barb 1 transformation event” and exhibiting fluorescence, using a first transgenic barb defined by the deposited sperm and a second barb that may be non-transgenic or transgenic fluorescent.
Breeding transgenic barbs carrying the “Red barb 1 transformation event”
Obtaining a first transgenic barb that exhibits fluorescence and comprises a chromosomally integrated expression cassette encoding a fluorescent protein, the expression cassette comprised by the “Red barb 1 transformation event,” sperm comprising the Red barb 1 transformation event having been deposited as ECACC accession no. 16121901, and breeding the first transgenic barb with a second barb to obtain a third transgenic barb hybrid comprising the “Red barb 1 transformation event.”
Using a non-transgenic second barb to form the hybrid
The method further includes breeding the first transgenic barb with a second barb that is non-transgenic.
Using a transgenic fluorescent second barb with a defined “Red barb 1 transformation event” deposit
The second barb is a transgenic fluorescent barb that contains a chromosomally integrated expression cassette for a fluorescent protein, specifically the cassette of the “Red barb 1 transformation event,” using sperm deposited as ECACC accession no. 1612901.
Requiring fluorescence in the third transgenic hybrid
The third transgenic barb exhibits fluorescence.
Specifying heterozygosity of the first barb for the “Red barb 1 transformation event”
The first barb is heterozygous for the “Red barb 1 transformation event.”
Specifying homozygosity of the first barb for the “Red barb 1 transformation event”
The first barb is homozygous for the “Red barb 1 transformation event.”
The claims focus on breeding barbs to produce a fluorescent third transgenic hybrid that carries the “Red barb 1 transformation event.” Dependent claims further define the second barb as non-transgenic or transgenic fluorescent and specify heterozygous or homozygous status for the first barb.
Stated Advantages
Provides transgenic red fluorescent ornamental fish that exhibit fluorescence.
Documented Applications
Ornamental fish market.
Research tools including cell lineage/migration tracing.
Research tools including genetic mosaic experiments/marking.
Research tools including fish cancer models.
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