Engineering protein posttranslational modification by PACE/furin in transgenic non-human mammals
First Claim
1. A non-human transgenic mammal which produces a post-translationally modified first protein, wherein said mammal comprises somatic and germ cells having stably incorporated into the genome of said cells, a first exogenous polynucleotide encoding a first protein susceptible to modification by PACE/furin and a second exogenous polynucleotide encoding PACE/furin, each of said first and second exogenous polynucleotides operatively linked to a mammary epithelial cell specific promoter, and wherein said mammal expresses said first exogenous polynucleotide and said second exogenous polynucleotide in the mammary gland epithelial cells, wherein said PACE/furin post-translationally modifies said first protein, such that said modification results in a post-translationally modified first protein.
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Abstract
The invention relates to transgenic non-human multicellular organisms that contain polynucleotides for expressing proteins that alter posttranslational modification. In particular, the invention provides multiply-transgenic animals in which a first transgene encodes a first protein, a second transgene encodes a second protein, and expression of the second protein affects the posttranlational modification of the first protein in cells of said organism. Expression in preferred embodiments is in specific cells and the modified protein is secreted into a bodily fluid. The invention provides related methods, proteins and products. An example provides transgenic animals that express human Protein C and the processing protease PACE/furin in mammary glands and secrete both proteins into milk.
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Citations
27 Claims
- 1. A non-human transgenic mammal which produces a post-translationally modified first protein, wherein said mammal comprises somatic and germ cells having stably incorporated into the genome of said cells, a first exogenous polynucleotide encoding a first protein susceptible to modification by PACE/furin and a second exogenous polynucleotide encoding PACE/furin, each of said first and second exogenous polynucleotides operatively linked to a mammary epithelial cell specific promoter, and wherein said mammal expresses said first exogenous polynucleotide and said second exogenous polynucleotide in the mammary gland epithelial cells, wherein said PACE/furin post-translationally modifies said first protein, such that said modification results in a post-translationally modified first protein.
- 14. A method for producing a post-translationally modified first protein in a non-human transgenic mammal, wherein said mammal comprises somatic and germ cells having stably incorporated into the genome of said cells, a first exogenous polynucleotide encoding a first protein susceptible to modification by PACE/furin and a second exogenous polynucleotide encoding PACE/furin, each of said first and second exogenous polynucleotides operatively linked to a mammary epithelial cell specific promoter, wherein said method comprises expressing said first exogenous polynucleotide and said second exogenous polynucleotide in the mammary gland eptithelial cells, and wherein said PACE/furin post-translationally modifies said first protein, such that said modification results in a post-translationally modified first protein, and obtaining said post-translationally modified first protein from the milk of said mammal.
Specification