Recombinant constructs and transgenic fluorescent ornamental fish therefrom

Inventors

Blake, Alan • Crockett, Richard • Essner, Jeffrey • Hackett, Perry • Nasevicius, Aidas

Assignees

Glofish LLC

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 fluorescent progeny fish of a transgenic fluorescent zebrafish in which both the zebrafish and progeny exhibit fluorescence and comprise one of the named transformation events: the Red zebrafish 1 transformation event, the Green zebrafish 1 transformation event, or the Yellow zebrafish 1 transformation event. The document further identifies deposited biological materials associated with the respective transformation events as ECACC accession no. 06090403, ECACC accession no. 06090401 and ECACC accession no. 06090402.

The disclosed recombinant DNA constructs use reef coral fluorescent proteins including DsRed2, ZsGreen1 and ZsYellow1, integrated into transgenic zebrafish germlines. Expression cassettes are identified as SEQ ID NO:1-5, with promoter selection spanning ubiquitous housekeeping promoters and tissue-specific promoters, including muscle-specific promoters such as zMLC (zebrafish fast skeletal muscle myosin light chain) promoter.

The document explains that Golden zebrafish (slc24a5 mutation) are used to enhance ornamental visibility and defines a concept of strong visible fluorescence. It describes preferred fluorescence expression breadth including ubiquitous expression versus chromatophore-specific expression involving melanophores, xanthophores and iridophores, and links the named transformation events to ornamental marketing and research/biosensing applications.

Claims Coverage

Two independent claims are present. Across them, the claims are grounded on three named ECACC-deposited transformation events and define fluorescent progeny fish or a method of producing transgenic fluorescent fish by obtaining a transformation-event zebrafish and breeding it.

The claim scope is centered on fluorescence-exhibiting transgenic zebrafish and progeny that include one of the three named transformation events associated with specific ECACC accession deposits, and on producing such transgenic fluorescent fish by obtaining the transformation-event zebrafish and breeding to obtain progeny comprising the same transformation event.

Stated Advantages

Documented Applications