Vector for integration site independent gene expression in mamma

Details Vector for integration site independent gene expression in mamma Vector for integration site independent gene expression in mamma

1995-06-07

1998-06-23

Rories, Charles C. P.

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

Method of regulating cell metabolism or physiology

435 691, 435 703, 435 704, 4351721, 4351723, 4353201, 800 2, 514 44, C12N 510, C12N 1513, C12N 1567, C12N 1585

Patent

active

057704490

ABSTRACT:
A vector for the integration of a gene into the genetic material of a mammalian host cell such that the gene may be expressed by the host cell. The vector comprises a promoter and the gene and in an immunoglobulin dominant control region derived from the mouse .lambda. immunoglobulin gene locus capable of eliciting host cell-type restricted, integration site independent, copy number dependent expression of said gene. The DNasel super hypersensitive site exemplified are i) about 2.35 kb upstream of the CAP site of the rearranged .lambda..sub.1 gene, ii) about 2.5 kb upstream of the genomic V.lambda..sub.2 segment or iii) about 30 kb downstream of the rearranged .lambda..sub.1 gene. Mammalian host cells transformed with the vector are disclosed as are transgenic mammals transformed with the vector and a method of producing a polypeptide comprising culturing a transformed mammalian cell. A method of gene therapy comprising the steps of i) removing stem cells from the body of a mammal, ii) optionally killing stem cells remaining in the body, iii) transforming the removed stem cells with the vector containing a gene deficient or absent in the body, and iv) replacing the transformed stem cells in the body is also disclosed. Also disclosed is functional mouse immunoglobulin .lambda..sub.1 enhancer consisting of a DNA sequence comprising all or a functional part of the DNA sequence between the EcoRI site 3.8 kb downstream of the Xho I site in the rearranged mouse .lambda..sub.1 gene and the SnaBI site 10 kb downstream of this Xho I site. The functional mouse immunoglobulin .lambda..sub.1 enhancer may comprise all or a functional part of i) the 1.3 kb first HindIII to HindIII DNA fragment downstream of the EcoRI site 3.8 kb downstream of the Xho I site in the rearranged mouse .lambda..sub.1 gene, ii) the 3.3 kb HindIII to HindIII DNA fragment downstream of the EcoRI site 3.8 kb downstream of the Xho I site in the rearranged mouse .lambda..sub.1 gene and spanning the SnaBI site 10 kb downstream of this Xho I site.

REFERENCES:
I. Weissman et al. Scientific American 269 (3) (Sep. '94) 65-71.
G. van Assendelft et al. Cell 56 (Mar. 24, 1989) 969-77.
A. Hienhuis et al. Cancer 67 ('91) 2700-4.
N. Sutkowski et al. P.N.A.S. 91 (Sep. '94) 8875-9.
J. Sharpe et al. P.N.A.S. 90 (Dec. '93) 11262-6.
L. Madisen et al. Genes & Development 8 ('94) 2212-26.
D. Talbot et al. Nature 338:352-5 ('89).
K. Mercola et al. Science 208:1033-5 (80).
M. Clive et al. Nature 284 422-5 ('80).
Antoniou et al., ".beta.-Globin dominant control region interacts differently with distal and proximal promoter Elements", Genes & Develolpment 4:1007-1013 (1990).
Brinster et al., "Introns increase transcriptional efficiency in transgenic mice", Proc. Natl. Acad. Sci. USA, vol. 85, pp. 836-840. (Feb. 1988).
Carson et al., "A linkage map of the mouse immunoglobulin lambda light chain locus", Immunogenetics, 29:173-179, (1989).
Greaves et al., "Human CD2 3'-Flanking Sequences Confer High-Level, T Cell-Specific, Position-Independent Gene Expression in Transgenic Mice", Cell, vol. 56, pp. 979-986, (Mar. 24, 1989).
Grosveld et al., "Position-Independent, High-Level Expression of the Human .beta.-Globin Gene in Transgenic Mice", Cell, vol. 51, 975-985, (Dec. 24, 1987).
Hesse et al., "Regulated gene expression in transfected primary chicken erythrocytes", Proc. Natl., Acad. Sci. USA, vol. 83, pp. 4312-4316, (Jun. 1986).
Lang et al., "The structure of the human CD2 gene and its expression in transgenic mice", The EMBO Journal, vol. 7, No. 6, pp. 1675-1682, (1988).
Le thi Bich-Thuy et al., "An enhancer associated with the mouse immunoglobulin .lambda.1 gene is specific for .lambda.light chain producing cells," Nucleic Acids Research, vol. 17, No. 13, pp. 5307-5321, (1989).
Maniatis et al., "Regulation of Inducible and Tissue-Specific Gene Expression", Science, vol. 236, (Jun. 5, 1987).
McDougall et al., "A transcriptional Enhancer 3'C.sub..beta.2 in the T Cell Receptor .beta. Locus", Science, vol. 241, p. 205-208, (Jul. 8, 1988).
Palmiter et al., "Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes", Nature, vol. 300, (Dec. 16, 1982).
Picard et al., "A lymphocyte-specific enhancer in the mouse immunoglobulin K gene", Nature, vol. 307, pp. 80-82, (Jan. 5, 1985).
Stief et al., "A Nuclear DNA attachment element mediates elevated and position-independent gene activity", Nature, vol. 341, (Sep. 28, 1989).
Storb et al., "High expression of cloned immunoglobulin K gene in transgenic mice is restricted to B lymphocytes", Nature, vol. 310, pp. 238-241, (Jul. 19, 1994).

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