WO2006005064B1 - Detection de 5-methylcytosine, compositions et methodes associees - Google Patents
Detection de 5-methylcytosine, compositions et methodes associeesInfo
- Publication number
- WO2006005064B1 WO2006005064B1 PCT/US2005/023761 US2005023761W WO2006005064B1 WO 2006005064 B1 WO2006005064 B1 WO 2006005064B1 US 2005023761 W US2005023761 W US 2005023761W WO 2006005064 B1 WO2006005064 B1 WO 2006005064B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- methylcytosine
- discriminator
- moiety
- detecting
- compound according
- Prior art date
Links
- LRSASMSXMSNRBT-UHFFFAOYSA-N 5-methylcytosine Chemical compound CC1=CNC(=O)N=C1N LRSASMSXMSNRBT-UHFFFAOYSA-N 0.000 title claims abstract 79
- 238000000034 method Methods 0.000 title claims abstract 18
- 239000000203 mixture Substances 0.000 title claims abstract 7
- 238000001514 detection method Methods 0.000 title claims 6
- 239000002773 nucleotide Substances 0.000 claims abstract 16
- 125000003729 nucleotide group Chemical group 0.000 claims abstract 16
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract 10
- 239000001257 hydrogen Substances 0.000 claims abstract 10
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 claims abstract 8
- 108020004707 nucleic acids Proteins 0.000 claims abstract 6
- 102000039446 nucleic acids Human genes 0.000 claims abstract 6
- 150000007523 nucleic acids Chemical class 0.000 claims abstract 6
- 238000010348 incorporation Methods 0.000 claims abstract 5
- 230000000295 complement effect Effects 0.000 claims abstract 3
- XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 claims abstract 3
- 235000011180 diphosphates Nutrition 0.000 claims abstract 3
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 claims abstract 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 claims abstract 2
- 150000001875 compounds Chemical class 0.000 claims 44
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical group O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 claims 14
- 108091034117 Oligonucleotide Proteins 0.000 claims 8
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims 6
- HDRRAMINWIWTNU-NTSWFWBYSA-N [[(2s,5r)-5-(2-amino-6-oxo-3h-purin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical group C1=2NC(N)=NC(=O)C=2N=CN1[C@H]1CC[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HDRRAMINWIWTNU-NTSWFWBYSA-N 0.000 claims 5
- 230000005764 inhibitory process Effects 0.000 claims 5
- 125000000217 alkyl group Chemical group 0.000 claims 4
- 238000006664 bond formation reaction Methods 0.000 claims 4
- 108020004414 DNA Proteins 0.000 claims 3
- 102000053602 DNA Human genes 0.000 claims 3
- 241000553086 Sybra Species 0.000 claims 3
- 150000001408 amides Chemical class 0.000 claims 3
- 229960002685 biotin Drugs 0.000 claims 3
- 235000020958 biotin Nutrition 0.000 claims 3
- 239000011616 biotin Substances 0.000 claims 3
- ABZLKHKQJHEPAX-UHFFFAOYSA-N tetramethylrhodamine Chemical compound C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=CC=C1C([O-])=O ABZLKHKQJHEPAX-UHFFFAOYSA-N 0.000 claims 3
- MPLHNVLQVRSVEE-UHFFFAOYSA-N texas red Chemical compound [O-]S(=O)(=O)C1=CC(S(Cl)(=O)=O)=CC=C1C(C1=CC=2CCCN3CCCC(C=23)=C1O1)=C2C1=C(CCC1)C3=[N+]1CCCC3=C2 MPLHNVLQVRSVEE-UHFFFAOYSA-N 0.000 claims 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims 2
- 125000003545 alkoxy group Chemical group 0.000 claims 2
- 239000011203 carbon fibre reinforced carbon Substances 0.000 claims 2
- 229910052736 halogen Inorganic materials 0.000 claims 2
- 150000002367 halogens Chemical class 0.000 claims 2
- 125000001072 heteroaryl group Chemical group 0.000 claims 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 claims 2
- 229910052698 phosphorus Inorganic materials 0.000 claims 2
- 239000011574 phosphorus Substances 0.000 claims 2
- 125000001424 substituent group Chemical group 0.000 claims 2
- 229910052717 sulfur Inorganic materials 0.000 claims 2
- 239000011593 sulfur Substances 0.000 claims 2
- YKBGVTZYEHREMT-KVQBGUIXSA-N 2'-deoxyguanosine Chemical group C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](CO)O1 YKBGVTZYEHREMT-KVQBGUIXSA-N 0.000 claims 1
- OCLZPNCLRLDXJC-NTSWFWBYSA-N 2-amino-9-[(2r,5s)-5-(hydroxymethyl)oxolan-2-yl]-3h-purin-6-one Chemical group C1=2NC(N)=NC(=O)C=2N=CN1[C@H]1CC[C@@H](CO)O1 OCLZPNCLRLDXJC-NTSWFWBYSA-N 0.000 claims 1
- 102000007347 Apyrase Human genes 0.000 claims 1
- 108010007730 Apyrase Proteins 0.000 claims 1
- XKMLYUALXHKNFT-UUOKFMHZSA-N Guanosine-5'-triphosphate Chemical group C1=2NC(N)=NC(=O)C=2N=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XKMLYUALXHKNFT-UUOKFMHZSA-N 0.000 claims 1
- 108060001084 Luciferase Proteins 0.000 claims 1
- 239000005089 Luciferase Substances 0.000 claims 1
- 102000004523 Sulfate Adenylyltransferase Human genes 0.000 claims 1
- 108010022348 Sulfate adenylyltransferase Proteins 0.000 claims 1
- 150000001721 carbon Chemical group 0.000 claims 1
- 229910052799 carbon Inorganic materials 0.000 claims 1
- 125000005842 heteroatom Chemical group 0.000 claims 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims 1
- 238000004806 packaging method and process Methods 0.000 claims 1
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 abstract 2
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 abstract 2
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 abstract 1
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 abstract 1
- 229940104302 cytosine Drugs 0.000 abstract 1
- 229940029575 guanosine Drugs 0.000 abstract 1
Abstract
L'invention concerne des compositions et des méthodes permettant de détecter 5-méthylcytosine dans un acide nucléique. Cette invention a aussi trait à un discriminateur de 5-méthylcytosine qui constitue un déoxyribonucléosidetriphosphate contenant un groupe caractéristique s'appariant à la cytosine, tel qu'une guanosine, et un groupe caractéristique qui entrave la liaison d'hydrogène entre le groupe caractéristique s'appariant à la cytosine et 5-méthylcytosine. Ce discriminateur est capable de s'apparier par la base à une cytosine, mais non à 5-méthylcytosine. 5-méthylcytosine compris par un nucléotide cible peut être détectée dans une réaction, au moyen d'une polymérase d'ADN et d'une amorce hybridée directement adjacente au nucléotide cible. Dans la réaction, le pyrophosphate libéré suite à l'incorporation d'un dNTP complémentaire à un nucléotide cible est détecté. L'absence d'incorporation du discriminateur, mais l'incorporation d'un dGTP, peut indiquer que le nucléotide cible constitue 5-méthylcytosine.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US10/881,363 | 2004-06-30 | ||
| US10/881,363 US7399614B2 (en) | 2004-06-30 | 2004-06-30 | 5-methylcytosine detection, compositions and methods therefor |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| WO2006005064A2 WO2006005064A2 (fr) | 2006-01-12 |
| WO2006005064A3 WO2006005064A3 (fr) | 2006-02-23 |
| WO2006005064B1 true WO2006005064B1 (fr) | 2006-04-20 |
Family
ID=35345122
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US2005/023761 WO2006005064A2 (fr) | 2004-06-30 | 2005-06-30 | Detection de 5-methylcytosine, compositions et methodes associees |
Country Status (2)
| Country | Link |
|---|---|
| US (2) | US7399614B2 (fr) |
| WO (1) | WO2006005064A2 (fr) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010037001A2 (fr) | 2008-09-26 | 2010-04-01 | Immune Disease Institute, Inc. | Oxydation sélective de 5-méthylcytosine par des protéines de la famille tet |
| US8486630B2 (en) | 2008-11-07 | 2013-07-16 | Industrial Technology Research Institute | Methods for accurate sequence data and modified base position determination |
| WO2010068289A2 (fr) | 2008-12-11 | 2010-06-17 | Pacific Biosciences Of California, Inc. | Classification de matrices d'acides nucléiques |
| US20230148447A9 (en) | 2008-12-11 | 2023-05-11 | Pacific Biosciences Of California, Inc. | Classification of nucleic acid templates |
| US9175338B2 (en) | 2008-12-11 | 2015-11-03 | Pacific Biosciences Of California, Inc. | Methods for identifying nucleic acid modifications |
| US8896806B2 (en) * | 2008-12-29 | 2014-11-25 | Nikon Corporation | Exposure apparatus, exposure method, and device manufacturing method |
| US9611510B2 (en) | 2011-04-06 | 2017-04-04 | The University Of Chicago | Composition and methods related to modification of 5-methylcytosine (5-mC) |
| US9238836B2 (en) | 2012-03-30 | 2016-01-19 | Pacific Biosciences Of California, Inc. | Methods and compositions for sequencing modified nucleic acids |
| AU2013240166A1 (en) | 2012-03-30 | 2014-10-30 | Pacific Biosciences Of California, Inc. | Methods and composition for sequencing modified nucleic acids |
| US9175348B2 (en) | 2012-04-24 | 2015-11-03 | Pacific Biosciences Of California, Inc. | Identification of 5-methyl-C in nucleic acid templates |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1990001069A1 (fr) | 1988-07-20 | 1990-02-08 | Segev Diagnostics, Inc. | Procede d'amplification et de detection de sequences d'acide nucleique |
| US5210015A (en) * | 1990-08-06 | 1993-05-11 | Hoffman-La Roche Inc. | Homogeneous assay system using the nuclease activity of a nucleic acid polymerase |
| US5436149A (en) * | 1993-02-19 | 1995-07-25 | Barnes; Wayne M. | Thermostable DNA polymerase with enhanced thermostability and enhanced length and efficiency of primer extension |
| GB9620209D0 (en) * | 1996-09-27 | 1996-11-13 | Cemu Bioteknik Ab | Method of sequencing DNA |
| GB9626815D0 (en) * | 1996-12-23 | 1997-02-12 | Cemu Bioteknik Ab | Method of sequencing DNA |
| DE10029915B4 (de) * | 2000-06-19 | 2005-06-16 | Epigenomics Ag | Verfahren zum Nachweis von Cytosin-Methylierungen |
| AU2003900368A0 (en) * | 2003-01-24 | 2003-02-13 | Human Genetic Signatures Pty Ltd | Assay for nucleic acid molecules |
| DE10328813B4 (de) * | 2003-06-20 | 2006-04-13 | Epigenomics Ag | Verfahren zur Untersuchung von Cytosin-Methylierungen in DNA-Sequenzen mittels triplexbildender Oligomere |
-
2004
- 2004-06-30 US US10/881,363 patent/US7399614B2/en not_active Expired - Fee Related
-
2005
- 2005-06-30 WO PCT/US2005/023761 patent/WO2006005064A2/fr active Application Filing
-
2008
- 2008-06-10 US US12/136,272 patent/US7999091B2/en not_active Expired - Lifetime
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