Recombinant constructs and transgenic fluorescent ornamental fish therefrom
Inventors
Blake, Alan • Crockett, Richard • Essner, Jeffrey • Hackett, Perry • Nasevicius, Aidas
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-10743526-B2
Publication Date
2020-08-18
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 transgenic fluorescent fish in which multiple fluorescent-protein transgenes are present in the fish genome and visibly express fluorescent color. In one embodiment, a first transgene encoding a first fluorescent protein is controlled by a muscle specific promoter, while a second transgene encoding a second fluorescent protein is controlled by a ubiquitous fish promoter, and a third transgene encoding a third fluorescent protein is controlled by a ubiquitous fish promoter.
The fish is defined to exhibit strong visible fluorescence over 75% to 100% of the body, optionally excluding fins and eyes. The first and second transgenes encode different fluorescent proteins, and the fish visibly expresses a fluorescent color from each of the first and second transgenes.
In another embodiment, the transgenic fluorescent fish includes first, second, and third transgenes, with each of the first, second, and third transgenes under the control of a muscle specific promoter. The fluorescent fish visibly expresses a fluorescent color different from the color encoded by the first and second transgenes, while still meeting the strong visible fluorescence criterion over 75% to 100% of the body.
Claims Coverage
The partial content provides coverage for two independent claims. Both claims define transgenic fluorescent fish with multiple fluorescent-protein transgenes, visible fluorescence, and strong whole-body fluorescence over 75% to 100% of the body, with differences primarily in promoter usage and in the allowable SEQ ID sequences.
Multiple fluorescent-protein transgenes with muscle-specific and ubiquitous promoters
A transgenic fluorescent fish comprising in its genome a first transgene encoding a first fluorescent protein under the control of a muscle specific promoter, a second transgene encoding a second fluorescent protein under the control of a ubiquitous fish promoter, and a third transgene encoding a third fluorescent protein under the control of a ubiquitous fish promoter.
Different fluorescent proteins with visible expression and strong body fluorescence
The first and second transgene encode different fluorescent proteins and the fluorescent fish visibly expresses a fluorescent color from each of the first and second transgene, wherein the transgenic fluorescent fish exhibits a strong visible fluorescence over 75% to 100% of the body of said fish, optionally excluding fins and eyes.
SEQ ID-controlled fluorescent protein sequences
At least one of the first and second transgene comprises a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:5.
All muscle-specific promoters for three transgenes
A transgenic fluorescent fish comprising in its genome a first transgene encoding a first fluorescent protein under the control of a muscle specific promoter, a second transgene encoding a second fluorescent protein under the control of a muscle specific promoter, and a third transgene encoding a third fluorescent protein under the control of a muscle specific promoter.
Different fluorescent proteins with visible expression and strong body fluorescence
The first and second transgene encode different fluorescent proteins and the fluorescent fish visibly expresses a fluorescent color different from the color encoded by the first and second transgene, wherein the transgenic fluorescent fish exhibits a strong visible fluorescence over 75% to 100% of the body of said fish, optionally excluding fins and eyes.
Restricted SEQ ID selection for muscle-driven transgenes
At least one of the first and second transgene comprises a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:4.
Overall, claim coverage centers on transgenic fluorescent fish with multiple fluorescent-protein transgenes that are visibly expressed and characterized by strong visible fluorescence over 75% to 100% of the body, optionally excluding fins and eyes. The main distinction between the independent claims is promoter configuration: one uses muscle-specific control for the first transgene and ubiquitous fish promoter control for the second and third transgenes, while the other uses muscle-specific promoters for the first, second, and third transgenes. Both claims additionally restrict the fluorescent-protein coding sequences via SEQ ID selection and require the first and second transgenes to encode different fluorescent proteins with corresponding visible fluorescent colors.
Stated Advantages
Provides strong visible fluorescence over 75% to 100% of the body of the fish, optionally excluding fins and eyes.
Enables visibly expressing fluorescent color from each of the first and second transgenes while encoding different fluorescent proteins.
Uses specified reef coral fluorescent protein sequences selected by SEQ ID to produce visible fluorescent colors in transgenic fish.
Documented Applications
No documented applications found
Interested in licensing this patent?