Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
1997-03-27
2001-04-10
Whisenant, Ethan (Department: 1655)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C536S023100, C536S024300
Reexamination Certificate
active
06214544
ABSTRACT:
Throughout this application, various references are referred to by number within parentheses. Disclosures of these publications in their entireties are hereby incorporated by reference into this application to more fully describe the state of the art to which this invention pertains. Full bibliographic citation for these references may be found at the end of this application, preceding the claims.
BACKGROUND OF THE INVENTION
The carcinogenic process is complex and often involves changes in the expression of two contrasting genetic elements, i.e., positive acting oncogenes and negative acting anti-oncogenes (tumor suppressor genes) (for reviews see references
1
-
3
). Compounds displaying selective toxicity toward transformed cells overexpressing different classes of oncogenes could prove useful as potential antitumor agents and as reagents for identifying cellular targets susceptible to modification by transforming oncogenes.
Cancer is often a consequence of changes in the expression of a number of genes. These include, dominant-acting oncogenes, tumor suppressor genes, genes affecting cell cycle and genes affecting genomic stability. In the case of tumor suppressor genes, the ability to identify and isolate these elements have proven difficult often involving extensive gene mapping and technically complex and many times unsuccessful molecular approaches. Prior to the art described in this invention, no simple and efficient way of identifying and cloning tumor suppressor genes has been available. The currently described approach is simple and effective in directly identifying potentially novel human tumor suppressor genes and directly cloning these genes. The approach, termed inducible suppression cDNA cloning, is useful in identifying both oncogene specific suppressor genes and global oncogene-independent tumor suppressor genes.
Current knowledge of tumor suppressor genes indicate that they often function as negative regulators of cell growth. Inherent in this operational definition of a tumor suppressor gene is the obvious implication that expression of a tumor suppressor gene in a target cell may evoke a loss of proliferative ability. This possibility has been demonstrated directly by reintroducing cloned tumor suppressor genes through DNA-transfection into tumor cells, i.e., growth and oncogenicity are suppressed. The growth inhibitory effect of tumor suppressor genes has prevented the previous development of functional assays permitting isolation of cells expressing novel tumor suppressor genes (anti-oncogenes).
SUMMARY OF THE INVENTION
This invention provides a method of identifying a tumor suppressor gene of a cell(s) which comprises the following steps: a) obtaining cDNA from a normal cell(s); b) preparing a library from the cDNA of step (a), wherein the cDNA is under the control of an inducible expression control system which also carries a selectable gene; c) introducing the vector library into a population of cell(s) expressing a transformed phenotype; d) placing the introduced transformed cell(s) from step (c) in conditions permitting expression of the cDNA and an effective concentration of an appropriate selection agent to select the cell(s) expressing the selectable gene; e) identifying the cell(s) which express the normal phenotype; and f) analyzing the cell(s) so identified so as to characterize the DNA and thus identify the tumor suppressor gene.
Analogous methods to identify tumor suppressors in normal cells and to identify genes associated with unknown genetic defects are also described.
REFERENCES:
patent: 5217889 (1993-06-01), Roninson et al.
patent: 5811234 (1998-09-01), Roninson et al.
Chen et al., Molecular and Cellular Biology 7(8): 2745-2752 (1987).*
Davis et al., Basic Methods in Molecular Biology, pp.296-297, Elsevier Science Publishing Co., New York, New York (1986).*
Deiss et al., Science 252 :117-120 (1991).*
Holzmayer et al., Nucleic Acids Research 20 :711-717 (1992).*
Artelt, P. et al., (1991) The prokaryotic neomycin-resistance-encoding gene acts as a transcriptional silencer in eukaryotic cells.Gene, 99:249-254.
Babiss, L.E. et al., (1986) Mutations in the E1a gene of adenovirus type 5 alter the tumorigenic properties of transformed cloned rat embryo fibroblast cells.Proc. Natl. Acad. of Sci., 83:2167-2171.
Bishop, J.M., (1987) The molecular genetics of cancer.Science, 235:305-311.
Boylon, J.F. et al., (1990) Role of the Ha-ras (rasH) oncogene in mediating progression of the tumor cell phenotype.Anticancer Research, 10:717-724.
Boylon, J.F. et al., (1992) Induction and progression of the transformed phenotype in cloned rat embryo fibroblast cells: studies employing type 5 adenovirus and wild-type and mutant Ha-ras oncogenes.Mol. Carcinogenesis, 5:118-128.
Dreher, K.L. and Cowan, K., (1991) Expression of antisense transcripts encoding an extracellular matrix protein by stably transfected vascular smooth muscle cells.European J. of Cell Biol., 54:1-9.
Fisher, P.B. et al., (1982) Analysis of type 5 adenovirus transformation with a cloned rat embryo line (CREF).Proc. Natl. Acad. of Sci., 79:3527-3531.
Fisher, P.B. et al., (1984) Enhancement of viral transformation and expression of the transformed phenotype by tumor promoters. In: Tumor Promotion and Carcinogenesis In Vitro, Mechanisms of Tumor Promotion, T.J. Slaga (ed.), Florida, CRC Press, pp. 57-123.
Fisher, P.B. and Rowley, P.T., (1991) Regulation of growth, differentiation and antigen expression in human tumor cells by recombiant cytokines: Application for the differentiation therapy of human cancerThe Status of Differentiation Therapy of Cancer, S. Waxman et al. (Eds.), New York, Raven Press, pp. 201-213.
Frenkel, K. et al., (1993) Inhibition of tumor promoter-mediated processes in mouse skin and bovine lens by caffeic acid phenethyl ester,Cancer Research, 53:1255-1261.
Glover, D. et al., (1989) Discloses that K-ras and Ki-ras are equivalent. “Oncogenes”,Oxford University Press, p. 217.
Gudkov, A.V. et al., (1994) Cloning mammalian genes by expression selection of genetic suppressor elements: Association of Kinesin with drug resistance and cell immortalization.Proc. Natl. Acad. of Sci. USA, 91:3744-3748.
Huang et al., (1988) Suppression of the neoplastic phenotype by replacement of the RB gene in human cancer cells.Science, 242:1563-1566.
Hu, M.C.-T. and Davidson, N., (1990) A combination of depression of the lac operator-repressor system with positive induction by glucocorticoid and metal ions provides a high-level-inducible gene expression system based on the human metallothionein-IIApromoter.Mol. and Cellular Biol., 10;6141-6151.
Johansen, T.E. et al., (1990) Biosynthesis of peptide precursors and protease inhibitors using new constitutive and inducible eukaryotic expression vectors.FEBS, 267:289-294.
Kitayama, H. et al., (1989) A ras-related gene with transformation suppressor activity.Cell, 56:77-84.
Lee et al., (1981) Glucocorticoids regulate expression of dihydrofolate reductase cDNA in mouse mammary tomour virus chimaeric plasmids,Nature, 294:228-232.
Levine, A.J. et al., (1993) The tumor suppressor genes.Ann. Rev. of Biochem., 62:623-651.
Lin, J. et al., (1994) Expression of the transformed phenotype induced by diverse acting viral oncogenes mediates sensitivity to growth suppression induced by caffeic acid phenethyl ester (CAPE).Intl. J. of Oncology, 4:1-11.
Liotta, L.A. et al., (1991) Review of cancer metastasis and angiogenesis: an imbalance of positive and negative regulation.Cell, 64:327-336.
Marshall, C.J. Tumor suppressor genes.Cell, 64:313-326.
von Melchner, H. et al., (1990) Isolation of cellular promoters by using a retrovirus promoter trap.Proc. Natl. Acad. of Sci., 87:3635-4022.
Noda, M. et al., (1983) Flat revertants isolated from Kirsten sarcoma virus-transformed cells are resistant to the action of specific oncogenes.Proc. Natl. Acad. of Sci., 80:5602-5606.
Noda, M. et al., (1989) Detection of genes with a potential for suppressing the transformed phenotype associated with activated ras genes.Proc. Natl. Acad. of Sci., 86:162-166.
Reid, L.H. et al., (1991) Cotra
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The Trustees of Columbia University
Whisenant Ethan
White John P.
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