Growth Differentiation Factor-8 Nucleic Acid and Polypeptide from Aquatic Species, and Transgenic Aquatic Species

Abstract

A transgenic non-human aquatic organisms, such as piscine, crustacean, mollusks, and the like, having a transgene which results in disrupting the production of and/or activity of growth differentiation factor-8 (GDF-8) chromosomally integrated into the germ cells of the aquatic organism is disclosed. Also disclosed are methods for making such organisms and nucleic acid sequences encoding GDF-8 polypeptides from such aquatic organisms.

1 . A transgenic non-human aquatic organism whose genome comprises a disruption of an endogenous growth differentiation factor-8 (GDF-8) gene, wherein said disruption comprises an insertion of a transgene, and wherein said disruption results in said animal exhibiting increased muscle mass as compared to wild-type animal. 
     
     
         2 . The transgenic aquatic organism of  claim 1 , wherein the aquatic organism is selected from the group consisting of a crustacean, a mollusk, a chordate, a gastropod, a pelecypod, a cephalopod and an echinoderm. 
     
     
         3 . The transgenic aquatic organism of  claim 1 , wherein the aquatic organism is selected from the group consisting of a piscine and an amphibian. 
     
     
         4 . The transgenic aquatic organism of  claim 1 , wherein the aquatic organism is homozygous or heterozygous for said disruption of an endogenous GDF-8 gene. 
     
     
         5 . Fish meat produced by the transgenic aquatic organism of  claim 3 . 
     
     
         6 . A method of producing food products from an aquatic organism having increased muscle mass, said method comprising:
 a) introducing a transgene that disrupts or interferes with expression of growth differentiation factor-8 (GDF-8) into a fertilized embryo, into a fertilized egg, or into germ cells of a pronuclear embryo of the aquatic organism   b) implanting an embryo of step a) into an oviduct of an appropriate pseudopregnant female, thereby allowing the embryo to mature to full term progeny;   c) testing the progeny for presence of the transgene to identify transgene-positive progeny having increased muscle mass;   d) cross-breeding transgene-positive progeny to obtain further transgene-positive progeny having increased muscle mass; and   e) processing the progeny having increased muscle mass to obtain food products.   
     
     
         7 . A method for producing a transgenic aquatic organism exhibiting an increase in muscle mass, said method comprising:
 a) introducing a transgene comprising a selectable marker sequence into a aquatic organism embryonic stem cell;   b) introducing said embryonic stem cell into an aquatic organism embryo;   c) transplanting said embryo into an appropriate pseudopregnant aquatic organism;   d) allowing said embryo to develop to term; and   e) identifying a transgenic aquatic organism whose genome comprises a disruption of the endogenous GDF-8 gene, wherein said disruption results in said aquatic organism exhibiting increased muscle mass as compared to a wild-type aquatic organism.   
     
     
         8 . A transgenic aquatic organism produced by the method of  claim 7 , wherein the genome of said aquatic organism comprises a disruption of an endogenous GDF-8 gene, wherein said disruption results in said aquatic organism exhibiting increased muscle mass as compared to a wild-type aquatic organism. 
     
     
         9 . The method of  claim 7 , wherein said transgenic aquatic organism is homozygous or heterozygous for said disruption of an endogenous GDF-8 gene. 
     
     
         10 . The method of  claim 7 , wherein said step of introducing a transgene into an embryonic stem cell is performed by infecting the embryonic stem cell with a virus containing the transgene. 
     
     
         11 . The method of  claim 10 , wherein the virus is a retrovirus. 
     
     
         12 . The transgenic aquatic organism of  claim 7 , wherein the aquatic organism is selected from the group consisting of a piscine and an amphibian. 
     
     
         13 . A method of producing aquatic organism food products having reduced cholesterol levels, said method comprising:
 a) introducing a transgene disrupting or interfering with expression of growth differentiation factor-8 (GDF-8) into an embryo of an aquatic organism;   b) culturing the embryo under conditions whereby progeny are hatched;   c) testing the progeny for presence of the transgene to identify transgene-positive progeny having reduced cholesterol levels;   d) cross-breeding transgene-positive progeny having reduced cholesterol levels; and   e) processing the progeny to obtain aquatic organism food products having reduced cholesterol levels.   
     
     
         14 . The method of  claim 13 , wherein the transgene comprises a GDF-8 antisense polynucleotide.