Novel card proteins involved in cell death regulation

Inactive Publication Date: 2004-12-16
THE BURNHAM INST
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Problems solved by technology

For example, a NAC which does not induce apoptosis may form hetero-oligomers with a NAC which is apoptotic, thus interfering with the apoptosis-inducing activity of NAC.
Although such polypeptides are considered NAPs or CAPs, it is well known that a cDNA sequence obtained f

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  • Novel card proteins involved in cell death regulation
  • Novel card proteins involved in cell death regulation
  • Novel card proteins involved in cell death regulation

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Embodiment Construction

[0181] cDNA Cloning.

[0182] Jurkat total RNA was reverse-transcribed to complementary DNAs using MMLV reverse transcriptase (Stratagene) and random hexanucleotide primers. Three overlapping cDNA fragments of NAC were amplified from the Jurkat complementary DNAs with Turbo P.function.u DNA polymerase (Stratagene) using the following oligonucleotide primer sets: primer set 1; 5'-CCGAATTCACCATGGCTGGCGGAGCCTGGGGC-3' (forward; SEQ ID NO:13) and 5'-CCGCTCGAGTCAACAGAGGGTTGTGGTGGTCTTG-3' (reverse; SEQ ID NO:14), primer set 2; 5'-CCCGAATTCGAACCTCGCATAGTCATACTGC-3' (forward; SEQ ID NO:15) and 5'-GTCCCACAACAGAATTCAATCTCAACGGTC-3' (reverse; SEQ ID NO:16), and primer set 3; 5'-TGTGATGAGAGAAGCGGTGAC-3' (forward; SEQ ID NO:17) and 5'-CCGCTCGAGCAAAGAAGGGTCAGCCAAAGC-3' (reverse; SEQ ID NO:18). The resultant cDNA fragments were ligated into mammalian expression vector pcDNA-myc (Invitrogen, modified as described in Roy et al., EMBO J. 16:6914-6925 (1997)) and assembled to full-length cDNA by ligating ...

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Abstract

The present invention provides NB-ARC and CARD-containing proteins (NACs), nucleic acid molecules encoding NACs and antibodies specific for at least one NAC. The invention further provides chimeric NAC proteins. The invention also provides screening assays for identifying an agent that can effectively alter the association of a NAC with a NAC-associated protein. The invention further provides methods of modulating apoptosis in a cell by introducing into the cell a nucleic acid molecule encoding a NAC or an antisense nucleotide sequence. The invention also provides a method of using a reagent that can specifically bind to a NAC to diagnose a pathology that is characterized by an increased or decreased level of apoptosis in a cell.

Description

[0001] This application is a divisional of U.S. Ser. No. 09 / 388,221, filed Sep. 1, 1999, which is incorporated herein by reference in its entirety.[0002] This invention relates generally to the fields of molecular biology and molecular medicine and more specifically to the identification of proteins involved in programmed cell death and associations of these proteins.BACKGROUND INFORMATION[0003] Programmed cell death is a physiologic process that ensures homeostasis is maintained between cell production and cell turnover in essentially all self-renewing tissues. In many cases, characteristic morphological changes, termed "apoptosis," occur in a dying cell. Since similar changes occur in different types of dying cells, cell death appears to proceed through a common pathway in different cell types.[0004] In addition to maintaining tissue homeostasis, apoptosis also occurs in response to a variety of external stimuli, including growth factor deprivation, alterations in calcium levels, ...

Claims

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Application Information

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IPC IPC(8): G01N33/53A61K31/7088A61K38/00A61K39/395A61K45/00A61K48/00A61P9/00A61P13/08A61P17/02A61P19/02A61P29/00A61P31/00A61P35/00A61P35/02A61P37/06A61P43/00C07K14/47C07K16/18C07K19/00C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12P21/08C12Q1/02C12Q1/68G01N33/566
CPCA01K2217/05A61K38/00A61K2039/505C07K14/4747C07K2319/00A61P9/00A61P13/08A61P17/02A61P19/02A61P29/00A61P31/00A61P35/00A61P35/02A61P37/06A61P43/00
Inventor REED, JOHN C.
Owner THE BURNHAM INST
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