CA2302134A1 - Composition and methods for monitoring the binding of a1/up1 to telomeric dna sequences and telomerase rna, and to measure the effect of this binding on telomere extension and protection - Google Patents
Composition and methods for monitoring the binding of a1/up1 to telomeric dna sequences and telomerase rna, and to measure the effect of this binding on telomere extension and protection Download PDFInfo
- Publication number
- CA2302134A1 CA2302134A1 CA002302134A CA2302134A CA2302134A1 CA 2302134 A1 CA2302134 A1 CA 2302134A1 CA 002302134 A CA002302134 A CA 002302134A CA 2302134 A CA2302134 A CA 2302134A CA 2302134 A1 CA2302134 A1 CA 2302134A1
- Authority
- CA
- Canada
- Prior art keywords
- binding
- sequence
- telomeric dna
- derivative
- telomerase rna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 108091035539 telomere Proteins 0.000 title claims abstract 10
- 210000003411 telomere Anatomy 0.000 title claims abstract 10
- 102000055501 telomere Human genes 0.000 title claims abstract 10
- 108010057210 telomerase RNA Proteins 0.000 title claims abstract 7
- 238000000034 method Methods 0.000 title claims 14
- 108091028043 Nucleic acid sequence Proteins 0.000 title claims 9
- 238000012544 monitoring process Methods 0.000 title 1
- 238000003556 assay Methods 0.000 claims abstract 7
- 108020004414 DNA Proteins 0.000 claims abstract 4
- 108010017842 Telomerase Proteins 0.000 claims abstract 2
- 239000012634 fragment Substances 0.000 claims 6
- 150000007523 nucleic acids Chemical class 0.000 claims 6
- 230000008436 biogenesis Effects 0.000 claims 5
- 239000003795 chemical substances by application Substances 0.000 claims 5
- 108020004707 nucleic acids Proteins 0.000 claims 3
- 102000039446 nucleic acids Human genes 0.000 claims 3
- 230000015572 biosynthetic process Effects 0.000 claims 2
- 230000001419 dependent effect Effects 0.000 claims 2
- 230000002255 enzymatic effect Effects 0.000 claims 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 claims 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 claims 1
- 239000000020 Nitrocellulose Substances 0.000 claims 1
- 101710163270 Nuclease Proteins 0.000 claims 1
- 108090000119 Nucleotidyltransferases Proteins 0.000 claims 1
- 102000003832 Nucleotidyltransferases Human genes 0.000 claims 1
- 108091028733 RNTP Proteins 0.000 claims 1
- 238000000338 in vitro Methods 0.000 claims 1
- 229920001220 nitrocellulos Polymers 0.000 claims 1
- 238000000159 protein binding assay Methods 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract 2
- 230000010076 replication Effects 0.000 abstract 2
- 108010014594 Heterogeneous Nuclear Ribonucleoprotein A1 Proteins 0.000 abstract 1
- 102000017013 Heterogeneous Nuclear Ribonucleoprotein A1 Human genes 0.000 abstract 1
- 230000000431 effect on proliferation Effects 0.000 abstract 1
- 230000009758 senescence Effects 0.000 abstract 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/136—Screening for pharmacological compounds
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Pathology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to the length of telomeres and to their effect on proliferation and senescence in cells. More specifically, it concerns the ability of hnRNP A1 and its shortened derivative UP1 to alter the length of telomeres in cells. More precisely, the invention relates to the ability of A1/UP1 to bind telomerase RNA, to bind and to protect mammalian telomeric DNA, and to modulate telomere extension and replication. Finally, the present invention relates to agents which can interfere with the binding of A1/UP1 to telomeres and telomerase, and to the use of protection, extension and replication assays to measure the biological impact of these agents.
Claims (13)
1. A method of identifying an agent which modulates telomere biogenesis in vitro comprising:
(a) incubating a nucleic acid target sequence for A1/UP1, wherein said target nucleic acid sequence is selected from telomerase RNA
and telomeric DNA, together with A1/UP1, a fragment thereof, or a derivative thereof, wherein said A1/UP1, a fragment thereof or derivative thereof is capable of binding to said target sequence; and (b) determining at least one of a binding between said A1/UP1, or fragment, or derivative thereof and said target nucleic acid sequence and an enzymatic activity dependent on the binding between said A1/UP1, or fragment, or derivative thereof and said target nucleic acid sequence;
wherein said agent is identified as a modulator of telomere biogenesis when said binding of A1/UP1, or fragment, or derivative thereof or said enzymatic activity is significantly different in the presence of said agent, as compared to in the absence thereof.
(a) incubating a nucleic acid target sequence for A1/UP1, wherein said target nucleic acid sequence is selected from telomerase RNA
and telomeric DNA, together with A1/UP1, a fragment thereof, or a derivative thereof, wherein said A1/UP1, a fragment thereof or derivative thereof is capable of binding to said target sequence; and (b) determining at least one of a binding between said A1/UP1, or fragment, or derivative thereof and said target nucleic acid sequence and an enzymatic activity dependent on the binding between said A1/UP1, or fragment, or derivative thereof and said target nucleic acid sequence;
wherein said agent is identified as a modulator of telomere biogenesis when said binding of A1/UP1, or fragment, or derivative thereof or said enzymatic activity is significantly different in the presence of said agent, as compared to in the absence thereof.
2. The method of claim 1, wherein said nucleic acid target sequence is telomeric DNA.
3. The method of claim 2, wherein binding between said A1/UP1 and said telomeric DNA is determined.
4. The method of claim 3, wherein said binding is determined by gel shift assay.
5. The method of claim 2, wherein a binding between said telomeric DNA sequence and said A1/UP1 is indirectly measured using a nuclease assay.
6. The method of claim 2, wherein a binding between said telomeric DNA sequence and said A1/UP1 is indirectly measured using a telomerase extension assay.
7. The method of claim 2, wherein a binding between said telomeric DNA sequence and said A1/UP1 is indirectly measured using a terminal nucleotidyl transferase (TdT) elongation assay.
8. The method of claim 2, wherein a binding between said telomeric DNA sequence and said A1/UP1 is indirectly measured using a rNTP-dependent DNA polymerase assay.
9. The method of claim 1, wherein said nucleic acid target sequence is telomerase RNA.
10. The method of claim 9, wherein a binding between said telomerase RNA sequence and said A1/UP1 is determined using a gel shift assay.
11. The method of claim 9, wherein a binding between said telomerase RNA sequence and said A1/UP1 is determined using a nitrocellulose binding assay.
12. A modulator of telomere biogenesis isolated using a method according to one of claims 1 to 11.
13. A method of identifying an agent which modulates telomere biogenesis comprising a determination of a formation of a ternary complex made up of:
(a) a telomeric DNA sequence;
(b) A1/UP1, or fragment thereof, or derivative thereof; and (c) telomerase RNA, wherein a modulator of telomere biogenesis is identified when a level of said formation of said ternary complex is detectably different in the presence of said agent as compared to in the absence thereof.
(a) a telomeric DNA sequence;
(b) A1/UP1, or fragment thereof, or derivative thereof; and (c) telomerase RNA, wherein a modulator of telomere biogenesis is identified when a level of said formation of said ternary complex is detectably different in the presence of said agent as compared to in the absence thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2302134A CA2302134C (en) | 1999-03-25 | 2000-03-27 | Composition and methods for monitoring the binding of a1/up1 to telomeric dna sequences and telomerase rna, and to measure the effect of this binding on telomere extension and protection |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA2,264,262 | 1999-03-25 | ||
| CA002264262A CA2264262A1 (en) | 1999-03-25 | 1999-03-25 | Composition and methods for modulating the length of telomeres |
| CA2302134A CA2302134C (en) | 1999-03-25 | 2000-03-27 | Composition and methods for monitoring the binding of a1/up1 to telomeric dna sequences and telomerase rna, and to measure the effect of this binding on telomere extension and protection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2302134A1 true CA2302134A1 (en) | 2000-09-25 |
| CA2302134C CA2302134C (en) | 2010-11-02 |
Family
ID=25680837
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2302134A Expired - Fee Related CA2302134C (en) | 1999-03-25 | 2000-03-27 | Composition and methods for monitoring the binding of a1/up1 to telomeric dna sequences and telomerase rna, and to measure the effect of this binding on telomere extension and protection |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2302134C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112326954A (en) * | 2020-10-28 | 2021-02-05 | 厦门宝太生物科技有限公司 | Homogeneous fluoroimmunoassay reagent for rapidly and quantitatively detecting D-dimer |
-
2000
- 2000-03-27 CA CA2302134A patent/CA2302134C/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112326954A (en) * | 2020-10-28 | 2021-02-05 | 厦门宝太生物科技有限公司 | Homogeneous fluoroimmunoassay reagent for rapidly and quantitatively detecting D-dimer |
| CN112326954B (en) * | 2020-10-28 | 2022-07-08 | 厦门宝太生物科技有限公司 | Homogeneous fluoroimmunoassay reagent for rapidly and quantitatively detecting D-dimer |
Also Published As
| Publication number | Publication date |
|---|---|
| CA2302134C (en) | 2010-11-02 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20180327 |
|
| MKLA | Lapsed |
Effective date: 20180327 |