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United States Patent | 5,733,746 |
Treco ,   et al. | March 31, 1998 |
The invention relates to novel human DNA sequences, targeting constructs, and methods for producing novel genes encoding thrombopoietin, DNase I, and .beta.-interferon by homologous recombination. The targeting constructs comprise at least: a) a targeting sequence; b) a regulatory sequence; c) an exon; and d) a splice-donor site. The targeting constructs, which can undergo homologous recombination with endogenous cellular sequences to generate a novel gene, are introduced into cells to produce homologously recombinant cells. The homologously recombinant cells are then maintained under conditions which will permit transcription of the novel gene and translation of the mRNA produced, resulting in production of either thrombopoietin, DNase I, or .beta.-interferon. The invention further relates to a methods of producing pharmaceutically useful preparations containing thrombopoietin, DNase I, or .beta.-interferon from homologously recombinant cells and methods of gene therapy comprising administering homologously recombinant cells producing thrombopoietin, DNase I, or .beta.-interferon to a patient for therapeutic purposes.
Inventors: | Treco; Douglas A. (Arlington, MA); Heartlein; Michael W. (Boxborough, MA); Hauge; Brian M. (Beverly, MA); Selden; Richard F. (Wellesley, MA) |
Assignee: | Transkaryotic Therapies, Inc. (Cambridge, MA) |
Appl. No.: | 406030 |
Filed: | March 17, 1995 |
Intern'l Class: | C12P 021/02 |
Field of Search: | 435/69.1,172.3,320.1,69.6 536/24.1,23.4,23.51 935/22,34,36,41,42,52,59,66 |
5272071 | Dec., 1993 | Chappel | 435/172. |
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__________________________________________________________________________ SEQUENCE LISTING (1) GENERAL INFORMATION: (iii) NUMBER OF SEQUENCES: 30 (2) INFORMATION FOR SEQ ID NO:1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 25 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1: AATTGCTCCTCGTGGTCATGCTTCT25 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: CTGTGAAGGACATGGGAGTCA21 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4488 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: TCTAGAGTCAGGATGGCACTGAAGGTCTCTGGGGAAGGGACGATGATGAGAGCCCGTCAG60 AAACCCTCCCCCCTTTCCTGGGTGATAGAGAAGACTCAGAACTTCACGCCCGGGGCTCTT120 TGCTCCCTACCTGCAGCCAGGGCCCGGTGCGATGAGAGCCCCCAGACCTCCCTGAAGGGT180 GAGTGAGTGTCACAAGTGCCACATGCAGCTGTTCTGCCCTAAGGAGCCGCAGAGACAACC240 GAGGCACTGCCCGCCACACCCCACAGACCTGGAGC0185 __________________________________________________________________________