TRANSGENIC RAINBOW SHARK

Abstract

The present invention relates to transgenic 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.

Claims

1.-80. (canceled)

81. A transgenic rainbow shark comprising a chromosomally integrated expression cassette encoding a fluorescent protein, wherein the rainbow shark comprises a transformation event selected from the group consisting of: the Orange rainbow shark 1 transformation event, sperm comprising the Orange rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010802); or the Purple rainbow shark 1 transformation event, sperm comprising the Purple rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010801); or the Blue rainbow 1 transformation event, sperm comprising the Blue rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010803); or the Green rainbow shark 1 transformation event, sperm comprising the Green rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010804).

82. The transgenic rainbow shark of claim 81, further defined as a fertile, transgenic rainbow shark.

83. The transgenic rainbow shark of claim 81, wherein the fish is homozygous for the integrated expression cassette.

84. The transgenic rainbow shark of claim 81, wherein the fish is heterozygous for the integrated expression cassette.

85. A method of providing a transgenic rainbow shark to the ornamental fish market, comprising obtaining a transgenic rainbow shark in accordance with claim 81, and distributing the fish to the ornamental fish market.

86. The method of claim 85, wherein the fish are distributed by a grower to a commercial distributor.

87. The method of claim 85, wherein the fish are distributed by a grower or a commercial distributor to a retailer.

88. The method of claim 87, wherein the retailer is a multi-product retailer having an ornamental fish department.

89. A method of producing a transgenic rainbow shark comprising: (a) obtaining a first rainbow shark that comprises a chromosomally integrated expression cassette encoding a fluorescent protein, wherein the rainbow shark comprises a transformation event selected from the group consisting of: the Orange rainbow shark 1 transformation event, sperm comprising the Orange rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010802); or the Purple rainbow shark 1 transformation event, sperm comprising the Purple rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010801); or the Blue rainbow 1 transformation event, sperm comprising the Blue rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010803); or the Green rainbow shark 1 transformation event, sperm comprising the Green rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010804), (b) breeding the first rainbow shark with a second rainbow shark to provide a third rainbow shark, wherein the third rainbow shark comprises the same transformation event of the first rainbow shark.

90. The method of claim 89, wherein the second rainbow shark is a non-transgenic rainbow shark.

91. A progeny fish of the transgenic rainbow shark of claim 81 that comprises a chromosomally integrated expression cassette encoding a fluorescent protein, wherein the rainbow shark and progeny exhibit fluorescence and comprise a transformation event selected from the group consisting of: the Orange rainbow shark 1 transformation event, sperm comprising the Orange rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010802); or the Purple rainbow shark 1 transformation event, sperm comprising the Purple rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010801); or the Blue rainbow 1 transformation event, sperm comprising the Blue rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010803); or the Green rainbow shark 1 transformation event, sperm comprising the Green rainbow shark 1 transformation event being deposited at the ECACC (Accession Number 19010804),

92. The progeny fish of claim 91, wherein the progeny is a fertile, transgenic rainbow shark.

93. The progeny fish of claim 91, wherein the progeny is homozygous for the integrated expression cassette.

94. The progeny fish of claim 91, wherein the progeny is heterozygous for the integrated expression cassette.

95. A method of providing a transgenic fish to the ornamental fish market, comprising obtaining a progeny in accordance with claim 91, and distributing the progeny to the ornamental fish market.

96. The method of claim 95, wherein the transgenic fish are distributed by a grower to a commercial distributor.

97. The method of claim 96, wherein the transgenic fish are distributed by a grower or a commercial distributor to a retailer.

98. The method of claim 97, wherein the retailer is a multi-product retailer having an ornamental fish department.

99. A method of producing a transgenic fish comprising: (a) obtaining a first transgenic fish in accordance with claim 91; and (b) breeding the first transgenic fish with a second fish to provide a third fish, wherein the third fish comprises the same transformation event of the first transgenic fish.

100. The method of claim 99, wherein the second fish is a non-transgenic fish.

Description

EXAMPLES

[0046] Certain embodiments of the invention are further described with reference to the following examples. These examples are intended to be merely illustrative of the invention and are not intended to limit or restrict the scope of the present invention in any way and should not be construed as providing conditions, parameters, reagents, or starting materials that must be utilized exclusively in order to practice the art of the present invention.

Example 1Orange Transgenic Rainbow Shark

[0047] Transgenic fish exhibiting an orange color are provided. The specific transgenic events embodied in these fish are designated the Orange rainbow shark 1 transformation event. Sperm from these fish may be used to fertilize rainbow shark eggs and thereby breed transgenic rainbow shark that comprise these specific transgenic integration events. Sperm from this line was deposited at the European Collection of Cell Cultures (ECACC (Accession Number 19010802)), Public Health England, CRYOSTORES, Bld. 17, Porton Down, Salisbury, SP4 OJG, United Kingdom, under the provisions of the Budapest Treaty as Orange rainbow shark 1.

Example 2Purple Transgenic Rainbow Shark

[0048] Transgenic fish exhibiting a purple color are provided. The specific transgenic events embodied in these fish are designated the Purple rainbow shark 1 transformation event. Sperm from these fish may be used to fertilize rainbow shark eggs and thereby breed transgenic rainbow shark that comprise these specific transgenic integration events. Sperm from this line was deposited at the European Collection of Cell Cultures (ECACC (Accession Number 19010801)), Public Health England, CRYOSTORES, Bld. 17, Porton Down, Salisbury, SP4 OJG, United Kingdom, under the provisions of the Budapest Treaty as Purple rainbow shark 1.

Example 3Blue Transgenic Rainbow Shark

[0049] Transgenic fish exhibiting a blue color are provided. The specific transgenic events embodied in these fish are designated the Blue rainbow shark 1 transformation event. Sperm from these fish may be used to fertilize rainbow shark eggs and thereby breed transgenic rainbow shark that comprise these specific transgenic integration events. Sperm from this line was deposited at the European Collection of Cell Cultures (ECACC (Accession Number 19010803)), Public Health England, CRYOSTORES, Bld. 17, Porton Down, Salisbury, SP4 OJG, United Kingdom, under the provisions of the Budapest Treaty as Blue rainbow shark 1.

Example 4Green Transgenic Rainbow Shark

[0050] Transgenic fish exhibiting a green color are provided. The specific transgenic events embodied in these fish are designated the Green rainbow shark 1 transformation event. Sperm from these fish may be used to fertilize rainbow shark eggs and thereby breed transgenic rainbow shark that comprise these specific transgenic integration events. Sperm from this line was deposited at the European Collection of Cell Cultures (ECACC (Accession Number 19010804)), Public Health England, CRYOSTORES, Bld. 17, Porton Down, Salisbury, SP4 OJG, United Kingdom, under the provisions of the Budapest Treaty as Green rainbow shark 1.

[0051] The fluorescent transgenic fish have use as ornamental fish in the market. Stably expressing transgenic lines can be developed by breeding a transgenic individual with a wild-type fish, mutant fish, or another transgenic fish. The desired transgenic fish can be distinguished from non-transgenic fish by observing the fish in white light, sunlight, ultraviolet light, blue light, or any other useful lighting condition that allows visualization of the orange, purple, green or blue color of the transgenic fish.

[0052] The fluorescent transgenic fish should also be valuable in the market for scientific research tools because they can be used for embryonic studies such as tracing cell lineage and cell migration. Additionally, these fish can be used to mark cells in genetic mosaic experiments and in fish cancer models.

[0053] All of the compositions and/or methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the compositions and/or methods and in the steps or in the sequence of steps of the methods described herein without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents that are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope, and concept of the invention as defined by the appended claims.