Recombinant constructs and transgenic fluorescent ornamental fish therefrom
Inventors
Blake, Alan • Crockett, Richard • Essner, Jeffrey • Hackett, Perry • Nasevicius, Aidas
Assignees
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Publication Number
US-12256719-B2
Publication Date
2025-03-25
Expiration Date
Abstract
The present invention relates to the method and use of reef coral fluorescent proteins in making transgenic red, green and yellow fluorescent zebrafish. Preferably, such fluorescent zebrafish are fertile and used to establish a population of transgenic zebrafish and to provide 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 a method of producing a transgenic, fluorescent fish having improved fluorescence. The method uses a first transgene encoding a fluorescent protein under the control of a ubiquitous fish promoter and a second transgene encoding the same fluorescent protein under the control of a muscle-specific fish promoter.
The transgenic, fluorescent fish are generated by chromosomal introduction of the first and second transgenes positioned on a single vector. The method includes selecting transgenic fish exhibiting strong visible fluorescence over 75% to 100% of the body, optionally excluding fins and eyes.
After selecting the transgenic fish, the method includes obtaining transgenic progeny of the selected fish, where the progeny exhibits fluorescence. The technology is exemplified for ornamental zebrafish with reef coral fluorescent proteins, and it includes maintaining fluorescence through breeding.
Claims Coverage
The document identifies one independent claim directed to a method of producing transgenic fluorescent fish with improved fluorescence. The independent claim is supported by dependent claims that refine promoter selection, fluorescent protein selection, and an ornamental-market distribution aspect.
Two transgenes on a single vector with ubiquitous and muscle-specific promoters
a first transgene encoding a fluorescent protein under the control of a ubiquitous fish promoter, and a second transgene encoding the same fluorescent protein under the control of a muscle-specific fish promoter, wherein said transgenic, fluorescent fish were generated by chromosomal introduction of said first and second transgenes positioned on a single vector
Selecting fish with strong visible fluorescence over most of the body
selecting a transgenic fish exhibiting strong visible fluorescence over 75% to 100% of the body of said fish, optionally excluding fins and eyes
Obtaining fluorescent transgenic progeny from selected fish
obtaining transgenic progeny of said selected transgenic fish, wherein the progeny exhibits fluorescence
Overall, the claim coverage centers on using two chromosomally introduced transgenes for the same fluorescent protein, one driven by a ubiquitous fish promoter and one driven by a muscle-specific fish promoter on a single vector, followed by selecting fish with strong visible fluorescence across 75% to 100% of the body and then obtaining fluorescent transgenic progeny from the selected fish.
Stated Advantages
Improved fluorescence.
Strong visible fluorescence over 75% to 100% of the body of the fish, optionally excluding fins and eyes.
Documented Applications
Ornamental fish market use case by providing fluorescent transgenic fish prepared according to the method.
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