US6849448B1
Expired / Lapsed
US
⏱️ 0.0 years remaining
Pigs and pig cells having an inactivated α 1,3-galactosyl transferase gene
Assignee: Hospira Adelaide Pty Ltd
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Filing: 1997-12-04
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Granted: 2005-02-01
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Expires: 2017-12-04
Abstract
Abstract Human pre-formed xenoantibodies play an important role in the hyperacute rejection response in human xenotransplantation. Disclosed are materials and methods for removing or neutralizing such antibodies. Also disclosed are materials and methods for reducing or eliminating the epitopes in the donor organs that are recognized by such antibodies. Such epitopes are formed as the result of activity by the enzyme α-1,3 galactosyltransferase. The porcine gene encoding α-1,3 galactosyltransferase is disclosed, as are materials and methods for inactivating (“knocking out”) the α-1,3 galactosyltransferase gene in mammalian cells and embryos. Included are nucleic acid constructs useful for inactivating the α-1,3 galactosyltransferase gene in a target cell. Also disclosed is a novel leukemia inhibitory factor (T-LIF) that is useful for maintenance of embryonic stem cells and primordial germ cells in culture.
Inventors
- 👤 Anthony J. F. d'Apice
- 👤 Martin J. Pearse
- 👤 Allan J. Robins
- 👤 Robert J. Crawford
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-08 03:05
CPC Classifications
A; A61; A61K; A61K31/00; A61K31/70; A61K31/7004; A01; A01K; A01K67/00; A01K67/027; A01K67/0275; A01K67/0276; C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/54; C07K16/00; C07K16/40; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/8509; C12N9/00; C12N9/10; C12N9/1048; C12N9/1051; A01K2217/00; A01K2217/07; A01K2217/075; A01K2227/00; A01K2227/10; A01K2227/105; A01K2227/108; A01K2267/00; A01K2267/02; A01K2267/025; A01K2267/03; A61K38/00; C12N2800/00; C12N2800/30
Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
65 words
A purified and isolated nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (1) nucleotides 91-1203 of the porcine nucleic acid sequence with SEQ ID NO: 7, (2) a sequence encoding a porcine polypeptide having α-1,3 galactosyltransferase activity and having the amino acid sequence of SEQ ID NO:10, and (3) a sequence complementary to the sequence of (1) or (2).
Claim 2
Dependent Claim
65 words
A purified and isolated nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (1) nucleotides 91-1203 of the porcine nucleic acid sequence with SEQ ID NO: 7, (2) a sequence encoding a porcine polypeptide having α-1,3 galactosyltransferase activity and having the amino acid sequence of SEQ ID NO:10, and (3) a sequence complementary to the sequence of (1) or (2).
Claim 3
Dependent Claim
73 words
A host cell comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (1) nucleotides 91-1203 of the porcine nucleic acid sequence with SEQ ID NO: 7, (2) a sequence encoding a porcine polypeptide having α-1,3 galactosyltransferase activity and having the amino acid sequence of SEQ ID NO:10, and (3) a sequence complementary to the sequence of (1) or (2), wherein the cell is in vitro.
Claim 4
Dependent Claim
73 words
A host cell comprising a recombinant nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (1) nucleotides 91-1203 of the porcine nucleic acid sequence with SEQ ID NO: 7, (2) a sequence encoding a porcine polypeptide having α-1,3 galactosyltransferase activity and having the amino acid sequence of SEQ ID NO:10, and (3) a sequence complementary to the sequence of (1) or (2), wherein the cell is in vitro.
Claim 5
Dependent Claim
88 words
A porcine α-1,3 galactosyltransferase encoded by a nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of (1) nucleotides 91-1203 of the porcine nucleic acid sequence with SEQ ID NO: 7, (2) a sequence encoding a porcine polypeptide having α-1,3 galactosyltransferase activity and having the amino acid sequence of SEQ ID NO:10, and (3) a sequence complementary to the sequence of (1) or (2), wherein the polypeptide is expressed from the cell of claim 2 and is encoded by the recombinant nucleic acid molecule.
Claim 6
Dependent Claim
53 words
A DNA construct comprising a disrupted porcine α-1,3 galactosyltransferase gene, wherein the disruption is by insertion of an exogenous sequence into said gene such that the disruption prevents expression of functional α-1,3 galactosyltransferase, wherein she gene, prior to disruption, encodes a porcine α-1,3 galactosyltransferase with an amino acid sequence of SEQ ID NO:10.
Claim 7
Dependent Claim
53 words
A DNA construct comprising a disrupted porcine α-1,3 galactosyltransferase gene, wherein the disruption is by insertion of an exogenous sequence into said gene such that the disruption prevents expression of functional α-1,3 galactosyltransferase, wherein she gene, prior to disruption, encodes a porcine α-1,3 galactosyltransferase with an amino acid sequence of SEQ ID NO:10.
Claim 8
Dependent Claim
27 words
The DNA construct of claim 4 , wherein said disruption is within exon 4, exon 7, exon 8, or exon 9 of the porcine α-1,3 galactosyltransferase gene.
Claim 9
Dependent Claim
15 words
The DNA construct of claim 4 , wherein said exogenous sequence is a selectable marker.
Claim 10
Dependent Claim
27 words
The DNA construct of claim 6 , wherein said selectable marker is selected from the group consisting of the neo R gene and the hyg R gene.
Claim 11
Dependent Claim
48 words
The DNA construct of claim 4 , wherein said exogenous sequence is flanked at its 5′ and 3′ ends by FRT DNA elements, and wherein stop codons have been inserted 3′ to the selectable marker for each of the three reading frames for the porcine α-1,3 galactosyltransferase gene.
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