Transgene expression in a avians

Abstract

A transgenic avian containing in its genome an exogenous nucleotide sequence which includes a promoter component and a vector with reduced promoter interference wherein the exogenous nucleotide sequence is integrated into the genome and the avian.

1 . A transgenic avian containing in its genome an exogenous nucleotide sequence comprising a promoter component and a SIN vector wherein the exogenous nucleotide sequence is integrated into the genome and the avian produces an exogenous protein which is deposited in a hard shell egg laid by the avian.  
     
     
         2 . The transgenic avian of  claim 1  wherein the promoter component is an oviduct specific promoter.  
     
     
         3 . The transgenic avian of  claim 1  wherein the avian is selected from the group consisting of a chicken, a turkey and a quail.  
     
     
         4 . The transgenic avian of  claim 1  wherein the promoter component is linked to a coding sequence exogenous to the avian.  
     
     
         5 . The transgenic avian of  claim 1  wherein the promoter component is an avian ovomucoid promoter component.  
     
     
         6 . The transgenic avian of  claim 1  wherein the promoter component is an avian ovalbumin promoter component.  
     
     
         7 . The transgenic avian of  claim 1  wherein the promoter component is an avian lysozyme promoter component.  
     
     
         8 . The transgenic avian of  claim 1  wherein the exogenous protein is a therapeutic protein.  
     
     
         9 . The transgenic avian of  claim 1  wherein the exogenous protein is a cytokine.  
     
     
         10 . The transgenic avian of  claim 1  wherein the exogenous protein is selected from the group consisting of erythropoietin, GM-CSF, interferon, fusion protein, CTLA4-Fc fusion protein, growth hormones, cytokines, structural, interferon, lysozyme, β-casein, albumin, α-1 antitrypsin, antithrombin III, collagen, factors VIII, IX, X (and the like), fibrinogen, lactoferrin, protein C, tissue-type plasminogen activator (tPA), somatotropin, and chymotryp sin, immunoglobulins, antibodies, immunotoxins, factor VIII, b-domain deleted factor VIII, factor VIIa, factor IX, anticoagulants; hirudin, alteplase, tpa, reteplase, tpa, tpa—3 of 5 domains deleted, insulin, insulin lispro, insulin aspart, insulin glargine, long-acting insulin analogs, glucagons, tsh, follitropin-beta, fsh, pdgh, inf-beta, inf-alpha 1, ifn-alpha 2, inf-beta, inf-beta 1b, ifn-beta 1a, ifn-gamma, ifn-gamma 1b, il-2, il-1 1, hbsag, ospa, dornase-alpha dnase, beta glucocerebrosidase, tnf-alpha, il-2-diptheria toxin fusion protein, tnfr-lgg fragment fusion protein laronidase, dnaases, alefacept, tositumomab, murine mab, alemtuzumab, rasburicase, agalsidase beta, teriparatide, parathyroid hormone derivatives, adalimumab (lggl), anakinra, biological modifier, nesiritide, human b-type natriuretic peptide (hbnp), colony stimulating factors, pegvisomant, human growth hormone receptor antagonist, recombinant activated protein c, omalizumab, immunoglobulin e (lge) blocker, lbritumomab tiuxetan, ACTH, glucagon, somatostatin, somatotropin, thymosin, parathyroid hormone, pigmentary hormones, somatomedin, luteinizing hormone, chorionic gonadotropin, hypothalmic releasing factors, etanercept, antidiuretic hormones, prolactin and thyroid stimulating hormone, an immunoglobulin polypeptide, immunoglobulin polypeptide D region, immunoglobulin polypeptide J region, immunoglobulin polypeptide C region, immunoglobulin light chain, immunoglobulin heavy chain, an immunoglobulin heavy chain variable region, an immunoglobulin light chain variable region and a linker peptide.  
     
     
         11 . The transgenic avian of  claim 1  wherein the retrovirus is selected from the group consisting of avian leukosis virus vector (ALV), a murine leukemia virus (MLV) retroviral vector, moloney murine leukemia Virus (MMLV) and a lentiviral vector.  
     
     
         12 . A transgenic avian comprising an oviduct cell which contains an exogenous nucleotide sequence comprising a promoter component linked to an exogenous coding sequence contained in an integrated SIN vector wherein the exogenous coding sequence is expressed in the oviduct cell and is secreted from the oviduct cell.  
     
     
         13 . The transgenic avian of  claim 12  wherein the avian is a chicken.  
     
     
         14 . The transgenic avian of  claim 12  wherein the oviduct cell is a tubular gland cell.  
     
     
         15 . The transgenic avian of  claim 12  wherein the promoter component is an avian ovomucoid promoter component.  
     
     
         16 . The transgenic avian of  claim 12  wherein the promoter component is an avian ovalbumin promoter component.  
     
     
         17 . The transgenic avian of  claim 12  wherein the promoter component is an avian lysozyme promoter component.  
     
     
         18 . A method of producing an exogenous protein comprising producing a transgenic avian having a nucleotide sequence in its genome comprising a vector which is at least one of a SIN vector and an SC negative vector wherein the nucleotide sequence comprises a promoter component linked to an exogenous coding sequence.  
     
     
         19 . The method of  claim 18  wherein the exogenous coding sequence encodes a human protein.  
     
     
         20 . The method of  claim 18  wherein the exogenous coding sequence encodes a therapeutic protein.  
     
     
         21 . The method of  claim 18  wherein the promoter component comprises a functional promoter sequence of a promoter selected from the group consisting of avian ovalbumin promoter component, avian ovomucoid promoter component, avian lysozyme promoter component and avian conalbumin promoter component.  
     
     
         22 . The method of  claim 18  wherein the avian is a chicken.  
     
     
         23 . A transgenic avian containing in its genome an exogenous nucleotide sequence comprising a promoter component and a SC negative vector wherein the exogenous nucleotide sequence is integrated into the genome and the avian produces an exogenous protein.  
     
     
         24 . The transgenic avian of  claim 23  wherein the promoter component comprises a functional promoter sequence of a promoter selected from the group consisting of avian ovalbumin promoter component, avian ovomucoid promoter component and avian lysozyme promoter component.  
     
     
         25 . A nucleic acid 90% identical to a nucleic acid molecule selected from the group consisting of nucleotide sequences that contain: 
 1. 3.5 kb OV fragment (includes DHS I, II & III) 
 5′ UTR-5′ portion (from Exon L)  
 5′ UTR-3′ portion (from Exon 1);  
   2. 3.5 kb OV fragment (includes DHS I, II & III) 
 5′ UTR-5′ portion (from Exon L)  
 Intron A  
 5′ UTR-3′ portion (from Exon 1)  
 3′ UTR;  
   3. 3.5 kb OV fragment (includes DHS I, II & III) 
 5′ UTR-5′ portion (from Exon L)  
 Intron A  
 5′ UTR-3′ portion (from Exon 1);  
   4. 3.5 kb OV fragment (includes DHS I, II & III) 
 5′ UTR-5′ portion (from Exon L)  
 5′ UTR-3′ portion (from Exon 1)  
 3′ UTR;  
   5. 3.5 kb OV fragment (includes DHS I, II & III) 
 5′ UTR-5′ portion (from Exon L)  
 Intron A  
 5′ UTR-3′ portion (from Exon 1)  
 3′ UTR/DHSA(bp 13576 to 15163 of  FIG. 8 );  
   6. 3.5 kb OV fragment (includes DHS I, II & III) 
 5′ UTR-5′ portion (from Exon L)  
 5′ UTR-3′ portion (from Exon 1)  
 3′ UTR/DHSA(bp 13576 to 15163 of  FIG. 8 );  
   7. 3.5 kb OV fragment (includes DHS I, II & III) 
 5′ UTR-5′ portion (from Exon L)  
 Intron A  
 5′ UTR-3′ portion (from Exon 1)  
 partial 3′ UTR  
 RRE;  
   8. ALV CTE 
 3.5 kb OV fragment (includes DHS I, II & III)  
 5′ UTR-5′ portion (from Exon L)  
 Intron A  
 5′ UTR-3′ portion (from Exon 1)  
 partial 3′ UTR;  
 wherein,  
   3.5 kb OV fragment (includes DHS I, II & III): Start: 3199 End: 6659 of  FIG. 8  (SEQ ID NO: 22);    5′ UTR-5′ portion (from Exon L): Start: 6659 End: 6705 of  FIG. 8  (SEQ ID NO: 22);    5′ UTR-3′ portion (from Exon 1): Start: 8295 End: 8311 of  FIG. 8  (SEQ ID NO: 22);    3′ UTR: Start: 13576 End: 14209 of  FIG. 8  (SEQ ID NO: 22);    partial 3′ UTR: Start 13576 End: 13996 of  FIG. 8  (SEQ ID NO: 22);    Intron A: Start: 6706 End: 8294 of  FIG. 8  (SEQ ID NO: 22);    Exon L: Start: 6659 End: 6705 of  FIG. 8  (SEQ ID NO: 22);    Exon 1: Start: 8295 End: 8478 of  FIG. 8  (SEQ ID NO: 22);    DHS III: Start: 3253 End: 3559 of  FIG. 8  (SEQ ID NO: 22);    DHS II: Start: 5629 End: 6009 of  FIG. 8  (SEQ ID NO: 22);    DHS I: Start: 6359 End: 6659 of  FIG. 8  (SEQ ID NO: 22); and    RRE: shown in  FIG. 9   a  (SEQ ID NO: 25)    ALV CTE shown in  FIG. 9   b  (SEQ ID NO: 26)