Genomic modification
The invention includes transchromosomal avians and transchromosomal avian cells and methods for the introduction of artificial chromosomes into the genome of avians and avian cells.
1 . A transchromosomic avian wherein the genome of the avian includes a transgene comprising greater than about 5,000 nucleotides.
2 . The transchromosomic avian of claim 1 wherein the genome of the avian includes a transgene comprising between about 5,000 and about 50,000,000 nucleotides.
3 . The transchromosomic avian of claim 1 wherein the avian is a G1 transchromosomic avian.
4 . The transchromosomic avian of claim 1 wherein the avian is a G2 transchromosomic avian.
5 . The transchromosomic avian of claim 1 wherein the avian is a germline transchromosomic avian.
6 . The transchromosomic avian of claim 1 wherein the avian is selected from the group consisting of chicken, quail and turkey.
7 . The transchromosomic avian of claim 1 wherein the artificial chromosome comprises a centromere.
8 . The transchromosomic avian of claim 7 wherein the centromere is selected from the group consisting of an insect centromere, a mammalian centromere and an avian centromere.
9 . The transchromosomic avian of claim 1 wherein the transgene includes a coding sequence for a pharmaceutical protein.
10 . The transchromosomic avian of claim 1 wherein the transgene includes a gene expression controlling region.
11 . The transchromosomic avian of claim 10 wherein the gene expression controlling region is operable in a cell of an oviduct.
12 . The transchromosomic avian of claim 10 wherein the gene expression controlling region includes a promoter selected from the group consisting of a lysozyme promoter, an ovomucin promoter, an ovomucoid promoter and an ovalbumin promoter.
13 . The transchromosomic avian of claim 1 wherein the avian lays an egg comprising a heterologous protein.
14 . The egg of claim 13 wherein the heterologous protein is present in an amount greater than about 0.1 μg per hard-shell egg.
15 . The egg of claim 13 wherein the heterologous protein is present in an amount in a range of between about 0.1 μg per hard-shell egg and about 1 gram per hard-shell egg.
16 . A transchromosomic avian wherein the avian lays an egg comprising a pharmaceutical protein.
17 . The egg of claim 16 wherein the pharmaceutical protein is present in egg white of the egg.
18 . The egg of claim 16 wherein the pharmaceutical protein is selected from the group consisting of a hormone, an antibody and a cytokine.
19 . The egg of claim 16 wherein the pharmaceutical protein is a cytokine.
20 . The egg of claim 16 wherein the cytokine is selected from the group consisting of GM-CSF, G-CSF, erythropoietin and interferon.
21 . The egg of claim 16 wherein the pharmaceutical protein is a monoclonal antibody.
22 . The egg of claim 16 wherein the pharmaceutical protein comprises a portion of an immunoglobulin.
23 . The egg of claim 13 wherein the heterologous protein is present in an amount greater than about 0.1 μg per milliliter of egg white.
24 . A transchromosomic avian wherein the avian lays an egg comprising a heterologous protein which is present in an amount greater than about 0.1 μg per hard-shell egg.
25 . The transchromosomic avian of claim 24 wherein the heterologous protein is present in an amount in a range of between about 0.1 μg per hard-shell egg and about 1 gram per hard-shell egg.
26 . The transchromosomic avian of claim 24 wherein the avian is a germline transchromosomic avian.
27 . The transchromosomic avian of claim 24 wherein the avian is selected from the group consisting of chicken, quail and turkey.
28 . The transchromosomic avian of claim 13 wherein the heterologous protein is a pharmaceutical protein.
29 . A method of dispersing nucleic acid in a cell comprising introducing nucleic acid into a cell in the presence of a cationic polymer in an amount sufficient to disperse the nucleic acid in the cell cytoplasm.