Production of Transgenic Avians Using Improved Retroviral Vectors
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 method of producing an exogenous protein comprising:
producing a transgenic avian whose genome comprises a self-inactivation (SIN) vector, wherein said vector comprising:
i) an avian leukosis virus (ALV)-based 5′ long terminal repeat (LTR); and
ii) a nucleic acid sequence encoding an exogenous protein operably linked to a promoter that causes expression in oviduct cells of the transgenic avian,
wherein said genome does not contain the nucleic acid sequence set forth from position 1 to position 173 of SEQ ID NO: 29 or a portion thereof, which inactivates transcription of a viral sequence in the vector, and wherein said transgenic avian produces the exogenous protein in egg white.
2 . The method of claim 1 , wherein the said ALV-based LTR comprises the nucleotide sequence position 174 to position 346 of SEQ ID NO: 29, or a nucleic acid sequence having at least 90% identity thereto.
3 . The method of claim 1 , wherein the exogenous coding sequence encodes a human protein.
4 . The method of claim 1 , wherein the exogenous coding sequence encodes a therapeutic protein.
5 . The method of claim 1 , wherein the promoter comprises a functional promoter sequence of a promoter selected from the group consisting of avian ovalbumin promoter, avian ovomucoid promoter, avian lysozyme promoter and avian conalbumin promoter.
6 . The method of claim 1 , wherein the avian is selected from the group consisting of a chicken, a turkey and a quail.
7 . The method of claim 1 , wherein the vector further comprises the nucleotide sequence set forth in SEQ ID NO: 25 or a nucleotide sequence having at least 90% identity thereto.
8 . The method of claim 7 , wherein the vector further comprises the nucleotide sequence set forth in SEQ ID NO: 26 or a nucleotide sequence having at least 90% identity thereto.
9 . The method of claim 1 , wherein said method further comprising isolating the exogenous protein from egg white of the avian.
10 . A transgenic avian whose genome comprises a self-inactivation (SIN) vector, said vector comprising:
i) an avian leukosis virus (ALV)-based 5′ long terminal repeat (LTR) having a deletion of the nucleic acid sequence set forth from position 1 to position 173 of SEQ ID NO: 29 or a portion thereof; and
ii) a nucleic acid sequence encoding an exogenous protein operably linked to a promoter that causes expression in oviduct cells of the transgenic avian,
wherein the deletion inactivates transcription activity of a viral sequence in the vector, and wherein said transgenic avian produces the exogenous protein in egg white.
11 . The transgenic avian of claim 10 , wherein the promoter comprises a functional promoter sequence of a promoter selected from the group consisting of avian ovalbumin promoter, avian ovomucoid promoter and avian lysozyme promoter.
12 . The transgenic avian of claim 10 , wherein the avian is selected from the group consisting of a chicken, a turkey and a quail.
13 . The transgenic avian of claim 10 , wherein the said ALV-based LTR comprises the nucleotide sequence position 174 to position 346 of SEQ ID NO: 29, or a nucleic acid sequence having at least 90% identity thereto.
14 . The transgenic avian of claim 10 , wherein the vector further comprises the nucleotide sequence set forth in SEQ ID NO: 25, or a nucleotide sequence having at least 90% identity thereto.
15 . The transgenic avian of claim 14 , wherein the vector further comprises the nucleotide sequence set forth in SEQ ID NO: 26, or a nucleotide sequence having at least 90% identity thereto.
16 . The transgenic avian of claim 10 , wherein the exogenous coding sequence encodes a human protein.
17 . The transgenic avian of claim 10 , wherein the exogenous coding sequence encodes a therapeutic protein.
18 . An egg laid by the transgenic avian of claim 10 .