Method of genetically altering and producing allergy free cats

Abstract

A transgenic cat with a phenotype characterized by the substantial absence of the major cat allergen, Fel d I. The phenotype is conferred in the transgenic cat by disrupting the coding sequence of the target gene with a specialized construct. The phenotype of the transgenic cat is transmissible to its offspring.

What is claimed is: 
     
         1 . An isolated nucleic acid comprising at least five contiguous nucleotides of the sequence depicted in SEQ ID NO. 7. 
     
     
         2 . A homologous recombination vector, comprising: (1) a first homology arm, (2) a desired polynucleotide, and (3) a second homology arm, wherein the desired polynucleotide is positioned between the first and second homology arms, and wherein each of the first and second homology arms comprises at least about a 1 kb sequence of SEQ ID NO. 7. 
     
     
         3 . The homologous recombination vector of  claim 2 , wherein the first homology arm comprises any sequence between nucleotide 1 to about nucleotide 8,800 of SEQ ID NO. 7. 
     
     
         4 . The homologous recombination vector of  claim 2 , wherein the first homology arm comprises any sequence between nucleotide 1 to about nucleotide 10,000 of SEQ ID NO. 7. 
     
     
         5 . The homologous recombination vector of  claim 2 , wherein the first homology arm comprises any sequence between nucleotide 1 to about nucleotide 10,800 of SEQ ID NO. 7. 
     
     
         6 . The homologous recombination vector of  claim 2 , wherein the first homology arm comprises any sequence between nucleotide 1 to about nucleotide 14,800 of SEQ ID NO. 7. 
     
     
         7 . The homologous recombination vector of  claim 2 , wherein the second homology arm comprises any sequence between about nucleotide 16,000 to nucleotide 21,939 of SEQ ID NO. 7. 
     
     
         8 . The homologous recombination vector of  claim 2 , wherein the second homology arm comprises any sequence between about nucleotide 14,700 to nucleotide 21,939 of SEQ ID NO. 7. 
     
     
         9 . The homologous recombination vector of  claim 2 , wherein the second homology arm comprises any sequence between about nucleotide 10,800 to nucleotide 21,939 of SEQ ID NO. 7. 
     
     
         10 . The homologous recombination vector of  claim 1 , wherein either or both of the homology arms comprises a SEQ ID NO. 7 sequence that has a length selected from the group consisting of about 1 kb long, about 2 kb long, about 3 kb long, about 4 kb long, about 5 kb long, about 6 kb long, about 7 kb long, about 8 kb long, about 9 kb long, and about 10 kb long. 
     
     
         11 . The homologous recombination vector of  claim 1 , wherein the desired polynucleotide is a selectable marker. 
     
     
         12 . A method for disrupting a target Fel d I sequence in a feline cell genome, comprising introducing the homologous recombination vector of  claim 1  into a feline cell, wherein (a) the homology arms of the vector function to recombine with the cell genome and (b) the desired polynucleotide integrates into the cell genome at the target sequence site thereby disrupting the target sequence. 
     
     
         13 . A feline cell, comprising the desired polynucleotide of  claim 12 .