Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of...
Reexamination Certificate
2011-08-30
2011-08-30
Falk, Anne-Marie (Department: 1632)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
C435S320100, C435S455000, C536S024100, C536S024310
Reexamination Certificate
active
08008071
ABSTRACT:
The subject invention pertains to nucleic acid constructs for post-transcriptional control of expression of a polynucleotide encoding a protein in a cell, wherein the constructs include a metabolite responsive instability element such as the glucose-regulated mRNA instability element. The subject invention further pertains to host cells and vectors comprising the nucleic acid constructs of the invention, as well as probes, methods, and kits for detecting metabolite responsive instability elements or mutations thereof. The present invention further concerns a reporter vector useful for detecting intracellular glucose and glucose-analogs, host cells genetically modified with the reporter vector, and methods for detecting intracellular glucose. The present invention utilizes an element that regulates messenger RNA (mRNA) stability in response to a metabolite such as glucose or a glucose analog. This glucose-regulated mRNA instability element has been mapped to the protein kinase C βII (PKCβII) mRNA that was found to decrease in the presence of elevated glucose levels. When cloned into a reporter vector, the region of PKCβII containing the mRNA instability element imparts glucose-sensitive instability to the mRNA that is transcribed, thereby down-regulating the expression of the reporter gene when glucose is elevated. The reporter vector of the present invention may be introduced into host cells, allowing detection of intracellular glucose and glucose analogs within intact, living cells in real-time and, optionally, in a high-throughput format.
REFERENCES:
patent: 5703054 (1997-12-01), Bennett et al.
patent: 5795961 (1998-08-01), Wallace et al.
patent: 6852529 (2005-02-01), Cooper et al.
GenBank Accession No. AA875851 (Mar. 25, 1998) alignment with SEQ ID No. 9.
Amara, F.M. et al. “Defining a Novelcis-Element in the 3′-Untranslated Region of Mammalian Ribonucleotide Reductase Component R2 mRNA”J. Biol. Chem.,Aug. 1996, 271(33):20126-20131.
Blobe, G.C. et al. “Protein Kinase C βII Specifically Binds to and Is Activated by F-actin”J. Biol. Chem.,Jun. 1996, 271(26):15823-15830.
Cañete-Soler, R. et al. “Characterization of Ribonucleoprotein Complexes and Their Binding Sites on the Neurofilament Light Subunit mRNA”J. Biol. Chem.,May 1998, 273(20):12655-12661.
Chalfant, C.E. et al. “Regulation of Alternative Splicing of Protein Kinase Cβ by Insulin”J. Biol. Chem.,Jun. 1995, 270(22):13326-13332.
Hara, K. et al. “1-Phosphatidylinositol 3-kinase activity is required for insulin-stimulated glucose transport but not for RAS activation in CHO cells”Proc. Natl. Acad. Sci.,USA, Aug. 1994, 91:7415-7419.
McClain, D.A. et al. “Glucose and glucosamine regulate growth factor gene expression in vascular smooth muscle cells”Proc. Natl. Acad. Sci.,USA, Sep. 1992, 89:8150-8154.
Nakshatri, H. and P. Bhat-Nakshatri “Multiple parameters determine the specificity of transcriptional response by nuclear receptors HNF-4, Arp-1, PPAR, RAR and RXR through common response elements”Nucleic Acids Res.,1998, 26(10):2491-2499.
Niino, Y.S. et al. “Positive and Negative Regulation of the Transcription of the Human Protein Kinase C β Gene”J. Biol. Chem.,Mar. 1992, 267(9):61513-6163.
Obeid, L.M. et al. “Cloning and Characterization of the Major Promoter of the Human Protein Kinase C β Gene”J. Biol. Chem.,Oct. 1992, 267(29):20804-20810.
Palmer, R.H. et al. “Activation of PRK1 by Phosphatidylinositol 4,5-Bisphosphate and Phosphatidylinositol 3,4,5-Trisphosphate”J. Biol. Chem.,Sep. 1995, 270(38):22412-22416.
Prokipcak, R.D. et al. “Purification and Properties of a Protein That Binds to the C-terminal Coding Region of Human c-mycmRNA”J. Biol. Chem.,Mar. 1994, 269(12):9261-9269.
Quilliam, L.A. et al. “Isolation of a NCK-associated Kinase, PRK2, an SH3-binding Protein and Potential Effector of Rho Protein Signaling”J. Biol. Chem.,Nov. 1996, 271(46):28772-28776.
Ron, D. et al. “Cloning of an intracellular receptor for protein kinase C: A homolog of the β subunit of G proteins”Proc. Natl. Acad. Sci. USA,Feb. 1994, 91:839-843.
Ross, J. “mRNA Stability in Mammalian Cells”Microbiology Reviews,Sep. 1995, 59(3):423-450.
Sayeski, P.P. and J.E. Kudlow “Glucose Metabolism to Glucosamine Is Necessary for Glucose Stimulation of Transforming Growth Factor-α Gene Transcription”J. Biol. Chem.,Jun. 1996, 271(25): 15237-15243.
Vincent, S. and Settleman, J. “The PRK2 Kinase Is a Potential Effector Target of both Rho and Rac GTPases and Regulates Actin Cytoskeletal Organization”Mol. Cell. Biol.,Apr. 1997, 17(4):2247-2256.
Amano, M. et al. “Identification of a Putative Target for Rho as the Serine-Threonine Kinase Protein Kinase N”Science,Feb. 1996, 271(5249):648-650.
Amara, F.M. et al. “Acis-transInteraction at the 3′-Untranslated Region of Ribonucleotide Reductase mRNA Is Regulated by TGF-β1,TGF-β2,and TGF-β3”Biochem. and Biophysical Res. Comm.,1996, 228:347-351.
Benbow, U. and C.E. Brinckerhoff “The AP-1 Site and MMP Gene Regulation: What Is All the Fuss About?”Matrix Biology,1997, 15:519-526.
Bernstein, P.L. et al. “Control of c-mycmRNA half-life in vitro by a protein capable of binding to a coding region stability determinant”Genes Dev.,1992, 6:642-654.
Chung, J. et al. “PDGF- and insulin-dependent pp70S6Kactivation mediated by phosphatidylinositol-3-OH kinase”Nature,Jul. 1994, 370:71-75.
Hansen, W.R. et al. “The 3′-nontranslated region of rat renal glutaminase mRNA contains a pH-responsive stability element”Amer. J. Physiology,Jul. 1996, 271:F126-F131.
Jia, Z. “Protein phosphatases: structures and implications”Biochem. Cell Biol,1997, 75:17-26.
Kemp, B.E. and R.B. Pearson “Protein kinase recognition sequence motifs”Trends Biochem. Sci.,Sep. 1990, 15(9):342-346.
Mukai, H. and Y. Ono “A Novel Protein Kinase with Leucine Zipper-Like Sequences: Its Catalytic Domain is Highly Homologous to That of Protein Kinase C”Biochem. Biophsy. Res. Comm.,Mar. 1994, 199(2):897-904.
Newton, a.C. “Regulation of protein kinase C”Curr. Opin. Cell Biol,1997, 9:161-167.
Nishizuka, Y. “Studies and Perspectives of Protein Kinase C”Science,Jul. 1986, 233(4761):305-312.
Nishizuka, Y. “Intracellular Signaling by Hydrolysis of Phospholipids and Activation of Protein Kinase C”Science,Oct. 1992, 258(5082):607-614.
Nishizuka, Y. “Protein kinase C and lipid signaling for sustained cellular responses”Faseb J.,1995, 9:484-496.
Orekhov, A.N. et al. “Cellular Composition of Atherosclerotic and Uninvolved Human Aortic Subendothelial Intima”Am J. Pathol.,1984, 115:17-24.
Palmer, R.H. and P.J. Parker “Expression, purification and characterization of the ubiquitous protein kinase C-related kinase 1”Biochem. J.,1995, 309:315-320.
Patel, N.A. et al. “Acute hyperglycemia regulates transcription and posttranscriptional stability of PKCβII mRNA in vascular smooth muscle cells”Faseb J.,1999, 13:103-113.
Perrotti, D. et al. “TLS/FUS, a pro-oncogene involved in multiple chromosomal translocations, is a novel regulator of BCR/ABL-mediated leukemogenesis”The EMBO J.,1998, 17(15):4442-4455.
Prekeris, R. et al. “Identification and Localization of an Actin-binding Motif That Is Unique to the Epsilon Isoform of Protein Kinase C and Participates in the Regulation of Synaptic Function”J. Cell Biol.,Jan. 1996, 132(1-2):77-90.
Ridley, A.J. et al. “The Small GTP-Binding Protein rac Regulates Growth Factor-Induced Membrane Ruffling”Cell,Aug. 1992, 70:401-410.
Ross, R. “The Pathogenesis of Atherosclerosis—An Update”N. Eng. J. Med.,Feb. 1986, 314(8):488-500.
Santen, R.J. et al. “Atheroscleros
Cooper Denise R.
Patel Niketa A.
Falk Anne-Marie
Saliwanchik Lloyd & Eisenschenk
University of South Florida
LandOfFree
Compositions and methods for detecting intracellular glucose... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Compositions and methods for detecting intracellular glucose..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Compositions and methods for detecting intracellular glucose... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2748477