Transkaryotic implantation
AssigneeShire Human Genetic Therapies, Inc.
InventorRichard F. Selden
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Abstract
The present invention provides a method of altering the concentration or level of expression of a desired gene product in a recipient subject which involves administering to a recipient a transkaryotic cell capable of expressing a desired gene when the cell is present within the subject. The invention further pertains to compounds produced by such cells, by transkaryotic animals in response to such cells, and to the uses of such compounds.
Claims15
A method of transferring a plasmid containing a DNA sequence coding for a protein into a recipient subject comprising:
- (a) transfecting autologous somatic cells in vitro with a plasmid containing a DNA sequence coding for a protein and which comprises a promoter by chemical or physical techniques to introduce the plasmid containing a DNA sequence coding for a protein into the cells;
- (b) screening the resulting transfected somatic cells in vitro to select a cell, wherein the selected cell is stably transfected with the plasmid containing a DNA sequence coding for a protein so that the selected cell has the permanent capacity to direct expression of the DNA sequence coding for a protein;
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- (c) cloning and expanding the selected somatic cell in vitro;
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- (d) injecting the resulting transfected, isolated, autologous, screened, cloned, and expanded somatic cells into the recipient subject.
A method of transferring a plasmid containing a DNA sequence coding for a protein into a recipient subject comprising:
- (a) providing autologous somatic cells;
- (b) transfecting the somatic cells in vitro with a plasmid containing a DNA sequence coding for a protein and further comprising a promoter capable of functioning in the somatic cells, wherein the somatic cells are stably transfected with the plasmid containing a DNA sequence coding for a protein so that the somatic cells have the permanent capacity to direct expression of the DNA sequence coding for a protein upon induction of the promoter;
- (c) screening the resulting transfected somatic cells in vitro to select a transfected somatic cell, wherein the screening comprises characterizing the transfected somatic cell with respect to expression and regulation of the DNA sequence coding for a protein by assaying for translation of mRNA into protein;
- (d) cloning and expanding, in vitro, the transfected and screened somatic cell selected in step (c) to form the 105–1010 transfected, screened, cloned, and expanded somatic cells,
- (e) combining the 105–1010 transfected, screened, cloned, and expanded somatic cells with a physiologically acceptable buffer or carrier;
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- (f) injecting the resulting transfected, isolated, autologous screened, cloned, and expanded cell preparation into the recipient subject.
Dependencies are inferred from the claim text.