Schizophrenia-related voltage-gated ion channel gene and protein

An amino acid and polynucleotide technology, which uses in-situ gene markers to screen CanIon channel modulators, treats or prevents various diseases or symptoms, and the field of polypeptides encoded by CanIon genes can solve the problem of difficult prediction of ion channels that are specific for therapeutic intervention. disease and other problems

Inactive Publication Date: 2006-05-24
SERONO GENETICS INST SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Thus, despite the involvement of ion channels in CNS diseases, it is difficult to predict which ion channels are effective targets for therapeutic intervention in a particular disease

Method used

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  • Schizophrenia-related voltage-gated ion channel gene and protein
  • Schizophrenia-related voltage-gated ion channel gene and protein
  • Schizophrenia-related voltage-gated ion channel gene and protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0833] Identification of biallelic marker-DNA extracts

[0834] Donors are unrelated and healthy. They represent enough diversity to represent a heterogeneous population in France. DNA was extracted from 100 individuals and tested for biallelic markers.

[0835] 30 ml of peripheral venous blood was obtained from each donor in the presence of EDTA. 2000rpm. Cells (sediment) were collected by centrifugation for 10 minutes. With lysate (50ml final volume: 10mM Tris pH7.6, 5mM MgCl 2 , 10mM NaCl) to lyse red blood cells. After resuspending the cell pellet in lysate, the solution is centrifuged (10 minutes, 2000 rpm.) as many times as necessary to remove residual erythrocytes in the supernatant.

[0836] Lyse the leukocyte pellet overnight at 42°C with 3.7ml of lysate containing the following components:

[0837] -3ml TE 10-2 (Tris-HCl 10mM, EDTA 2mM) / NaCl 0.4M

[0838] -200 μl SDS 10%

[0839] - 500 μl K-protease (2 mg K-protease dissolved in TE 10-2 / NaCl 0.4M)

[0840...

Embodiment 2

[0845] Identification of biallelic markers: amplification of genomic DNA by PCR

[0846] Amplification of the specific genomic sequence of the DNA sample of Example 1 was performed on the DNA pool obtained above. In addition, 50 individual samples were similarly amplified.

[0847] Perform PCR assays using the following protocol:

[0848] Final volume 25μl

[0849] DNA 2ng / μl

[0850] MgCl 2 2mM

[0851] dNTP (each) 200μM

[0852] Primer (each) 2.9ng / μl

[0853] Ampli Taq Gold DNA Polymerase 0.05 unit / μl

[0854] PCR buffer (10x=0.1M trisHCl pH8.30.5M KCl) 1x

[0855] Each pair of first primers was designed using the CanIon gene sequence information disclosed herein and OSP software (Hilier & Green, 1991). This first pair of primers is approximately 20 nucleotides in length and has the sequence disclosed in Table 1 under the columns labeled PU and RP.

[0856]

amplicon

In SEQ ID1 this expansion

Adder's l...

Embodiment 3

[0862] Identification of biallelic markers - testing and identification of polymorphisms on amplified genomic DNA

[0863] Sequencing of the amplified DNA obtained in Example 2 was performed on an ABI 377 sequencer. The sequence of the amplified product was determined using an automated dideoxy terminator sequencing reaction with a dye terminator cycle sequencing program. The sequencing reaction products were electrophoresed on a sequencing gel, and their sequences were determined using gel imaging analysis (ABI Prism DNA Sequencing Analysis software (version 2.1.2)).

[0864] The sequence data were further evaluated to detect the presence of biallelic markers in the amplified fragments. Polymorphism searches are based on overlapping peaks resulting from the presence of different bases at the same position above in the electrophoretic pattern.

[0865] Of the 17 fragments amplified, 18 biallelic markers were detected. The positions of these biallelic markers are shown in Ta...

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Abstract

The invention concerns the genomic DNA, cDNA, and polypeptide sequences of CanIon, a novel voltage gated ion channel protein. The invention also concerns biallelic markers of the CanIon gene. The CanIon gene may be used as a biological target for the treatment and diagnosis of schizophrenia, bipolar disorder, and other diseases and conditions.

Description

technical field [0001] The present invention relates to voltage-gated ion channel genes and proteins and their roles in diseases. The present invention relates to polynucleotides encoding CanIon polypeptides and regulatory regions located at the 5'- and 3'-ends of said coding region. The present invention also relates to polypeptides encoded by CanIon genes. The present invention also provides methods for screening CanIon channel modulators (such as antagonists) and methods for using such modulators to treat or prevent various diseases or symptoms. The invention also relates to antibodies specific for such polypeptides and thus useful as diagnostic reagents. The invention further includes biallelic markers of the CanIon gene useful in genetic analysis. Background technique [0002] Basic and clinical researchers enable them to conduct increasingly in-depth studies of brain and nervous system function in health and disease. Numerous neurobiological and pharmacological hyp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/027C12N15/12A61K45/00A61P9/00A61P25/00A61P25/18C07K14/47C07K14/705C07K16/28C12M1/00C12M1/34C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12N15/63C12P21/02C12Q1/02C12Q1/68G01N33/15G01N33/50G01N33/53G01N33/566G01N33/58G01N37/00
CPCA01K2217/05C07K14/705A61P25/00A61P25/18A61P25/28A61P9/00C12N15/11
Inventor D·科恩I·丘马科弗A·-M·西蒙H·阿布德雷希姆
Owner SERONO GENETICS INST SA
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