JPH08507202A - リボザイム増幅診断用薬 - Google Patents
リボザイム増幅診断用薬Info
- Publication number
- JPH08507202A JPH08507202A JP6514281A JP51428194A JPH08507202A JP H08507202 A JPH08507202 A JP H08507202A JP 6514281 A JP6514281 A JP 6514281A JP 51428194 A JP51428194 A JP 51428194A JP H08507202 A JPH08507202 A JP H08507202A
- Authority
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- Japan
- Prior art keywords
- ribozyme
- molecule
- nucleic acid
- target
- target nucleic
- 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.)
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
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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
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
- C12Q1/6823—Release of bound markers
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/12—Type of nucleic acid catalytic nucleic acids, e.g. ribozymes
- C12N2310/121—Hammerhead
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/30—Chemical structure
- C12N2310/35—Nature of the modification
- C12N2310/351—Conjugate
- C12N2310/3517—Marker; Tag
Abstract
Description
Claims (1)
- 【特許請求の範囲】 1.溶液中の標的核酸分子を検出する方法であって、以下の工程: 溶液中に、2つの相補的なヌクレオチド分子がハイブリダイズし得る条件下で :リボザイム分子、標的核酸分子、およびリボザイムに対する開裂部位を含む標 識された共標的核酸分子、を提供する工程、ここで、該共標的分子および標的分 子は異なる配列を有し、そして該リボザイム分子は、該標識された共標的核酸分 子に相補的な領域および該標的核酸分子に相補的な領域を含み、ここで、該相補 的な領域は、少なくとも、該リボザイム分子と共標的核酸分子と標的核酸分子と の間の結合を得るために最小限の数の相補的ヌクレオチドを含み、 該リボザイム分子を、該標識された共標的核酸分子および標的核酸分子と反応 させる工程、および 該標的核酸分子が溶液中に存在する場合、該標的核酸分子が溶液中に存在しな い場合と比較して遊離の標識の存在を検出する工程: を包含する方法。 2.前記リボザイムが、テトラヒメナリボザイム、RNAase P、イモリのサテライ トリボザイム由来のリボザイム、植物ウイロイド由来のハンマーヘッドリボザイ ム、およびデルタ肝炎ウイルス由来のアクスヘッド(axehead)リボザイムから なる群 に由来する、請求項1に記載の方法。 3.前記標的核酸分子が共標的の第1の核酸分子により結合される、請求項1に 記載の方法。 4.前記標的核酸分子がタンパク質をコードする、請求項3に記載の方法。 5.前記標識された共標的核酸分子が固体支持体上に固定化されている、請求項 1に記載の方法。 6.前記標識された共標的核酸分子が、染料、発色性基質と反応性の酵素、蛍光 標識、化学発光標識、および放射性標識からなる群から選択される標識で標識さ れる、請求項1に記載の方法。 7.前記リボザイムが、該リボザイムが前記標的核酸分子に結合するときに、構 造的に活性である、請求項1に記載の方法。 8.前記リボザイムが、該リボザイムが前記標的核酸分子および標識された共標 的核酸分子の両者に結合するときに、構造的に活性である、請求項1に記載の方 法。 9.前記リボザイムの酵素活性を変える分子に対する結合に基づいて該リボザイ ム分子を選択する工程をさらに包含する、請求項1に記載の方法。 10.請求項9に記載の方法であって、さらに前記リボザイム分子を繰り返し合 成する工程;該リボザイム分子と、活性を変える分子とを、いくつかのリボザイ ム分子が該分子に結合し得る条件下で反応させる工程; 該活性を変える分子と結合しないリボザイムを除去する工程、 該リボザイムの活性について結合分子の影響をスクリーニングする工程、 該活性を変える分子に結合し、そしてそれによってその活性が変化し得るリボ ザイム分子を単離する工程、および 該分子と結合しそしてその活性が該分子との結合により変化し得るリボザイム 分子が単離されるまで、このプロセスを繰り返す工程;を包含する方法。 11.前記リボザイムが37℃を超える高温で安定で、そして標的核酸分子が37℃ を超える条件下で結合する、請求項1に記載の方法。 12.前記標的核酸分子が前記リボザイム分子により結合されるときに、該標的 核酸分子が結合されない場合に比べ検出 される標識がより少ない、請求項1に記載の方法。 13.前記標的核酸分子が前記リボザイム分子により結合されるときに、該標的 核酸分子が結合されない場合に比べ検出される標識がより多い、請求項1に記載 の方法。 14.リボザイム分子、標的核酸分子、および該リボザイムに対する開裂部位を 含む標識された共標的核酸分子を含む組成物であって、ここで、該共標的分子お よび標的分子は異なる配列を有し、そして該リボザイム分子は、該標識された共 標的核酸分子に相補的な領域および該標的核酸分子に相補的な領域を含み、ここ で、該相補的な領域は、少なくとも、該リボザイム分子と共標的核酸分子と標的 核酸分子との間の結合を得るために最小限の数の相補的ヌクレオチドを含む、組 成物。 15.前記共標的核酸分子が、標的核酸分子に結合し、そして前記リボザイム分 子により結合されおよび開裂される、請求項14に記載の組成物。 16.前記標的核酸分子がタンパク質をコードする、請求項14に記載の組成物 。 17.前記リボザイムが、テトラヒメナリボザイム、RNAase P、イモリのサテライトリボザイム由来のリボザイム、植物ウイロイド由来のハ ンマーへッドリボザイム、およびデルタ肝炎ウイルス由来のアクスヘッドリボザ イムからなる群に由来する、請求項14に記載の組成物。 18.前記リボザイムが、前記共標的核酸分子上の第1の部分に結合するときに 、構造的に活性である、請求項14に記載の組成物。 19.前記リボザイムが、第1の相補的な領域が共標的分子に結合し、そして第 2の相補的な領域が標的分子上の第2の部分に結合するときに、構造的に活性で ある、請求項14に記載の組成物。 20.前記リボザイムの酵素的活性が、該リボザイムへの制御性分子の結合によ り変えられる、請求項14に記載の組成物。 21.前記リボザイム分子が、37℃と60℃との間の温度で構造的に活性である、 請求項14に記載の組成物。 22.前記標識された共標的核酸分子が、固体支持体上に固定化されている、請 求項14に記載の組成物。 23.前記標識された共標的核酸が、染料、発色性基質と反応性の酵素、蛍光標 識、化学発光標識、および放射性標識からなる群から選択される標識で標識され る、請求項14に記載の組成物。 24.前記標的核酸分子が、前記リボザイムまたは共標的分子のいずれかにハイ ブリダイズする場合、該標的核酸分子が結合されない場合に比べ検出される標識 がより少ない、請求項14に記載の組成物。 25.前記標識核酸が、前記リボザイムまたは第2の分子のいずれかにハイブリ ダイズする場合、該標識核酸が結合されない場合に比べ検出される標識がより多 い、請求項14に記載の組成物。 26.リボザイム分子を選択する方法であって、該リボザイム活性が制御性分子 との結合により変えられ得、以下の工程; 該リボザイム分子を繰り返し合成する工程、 該リボザイム分子と、活性を変える分子とを、いくつかのリボザイム分子が該 分子に結合し得る条件下で反応させる工程、 該活性を変える分子と結合しないリボザイムを除去する工程、 該リボザイムの活性に関して結合分子の影響をスクリーニ ングし、該リボザイム分子を単離する工程、ここで、該分子が該リボザイムを結 合しそしてその活性に影響を与え、および 該分子と結合し、そしてその活性が該分子との結合により変えられ得るリボザ イム分子が単離されるまで、このプロセスを繰り返す工程、を包含する方法。 27.トランスに作用するリボザイム診断システムであって、染料、発色性基質 と反応性の酵素、蛍光標識、化学発光標識、および放射性標識からなる群から選 択される標識で標識され、そして標的分子に相補的な2つの領域を有するリボザ イムを含み、ここで、該標識は、該リボザイムが該標的分子に結合する場合、該 リボザイムから開裂される、システム。 28.前記標識リボザイムが、テトラヒメナリボザイム、RNAase P、イモリのサ テライトリボザイム由来のリボザイム、植物ウイロイド由来のハンマーヘッドリ ボザイム、およびデルタ肝炎ウイルス由来のアクスヘッドリボザイムからなる群 に由来する、請求項27に記載のリボザイム診断システム。 29.溶液中の標的された核酸分子を検出する方法であって、 トランスに作用するリボザイム診断システムを提供する工程であって、該シス テムは、染料、発色性基質と反応性の酵素、蛍光標識、化学発光標識、および放 射性標識からなる群 から選択される標識で標識され、そして該標的された分子に相補的な2つの領域 を有するリボザイムを含み、ここで、該標識は、該リボザイムが該標的された分 子に結合するときに開裂され;および 該リボザイムから開裂された該標識を検出することにより該標的された分子を 検出する工程、を包含する方法。 30.溶液中の標的された核酸分子を検出する方法であって、前記リボザイムが 、テトラヒメナリボザイム、RNAase P、イモリのサテライトリボザイム由来のリ ボザイム、植物ウイロイド由来のハンマーヘッドリボザイム、およびデルタ肝炎 ウイルス由来のアクスヘッドリボザイムからなる群に由来する、請求項29に記 載の方法。 31.染料、発色性基質と反応性の酵素、蛍光標識、化学発光標識、および放射 性標識からなる群から選択される標識で標識され、そして標的分子に相補的な2 つの領域を有し、ここで、該標識は、該リボザイムが該標的分子に結合するとき に開裂される、リボザイム 32.テトラヒメナリボザイム、RNAase P、イモリのサテライトリボザイム由来 のリボザイム、植物ウイロイド由来のハンマーヘッドリボザイム、およびデルタ 肝炎ウイルス由来のアクスヘッドリボザイムからなる群に由来する、請求項31 に記載のリボザイム。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US98530892A | 1992-12-04 | 1992-12-04 | |
US985,308 | 1992-12-04 | ||
PCT/US1993/011775 WO1994013833A1 (en) | 1992-12-04 | 1993-12-03 | Ribozyme amplified diagnostics |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2003310777A Division JP2004000281A (ja) | 1992-12-04 | 2003-09-02 | リボザイム増幅診断用薬 |
Publications (2)
Publication Number | Publication Date |
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JPH08507202A true JPH08507202A (ja) | 1996-08-06 |
JP3595841B2 JP3595841B2 (ja) | 2004-12-02 |
Family
ID=25531365
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP51428194A Expired - Lifetime JP3595841B2 (ja) | 1992-12-04 | 1993-12-03 | リボザイム増幅診断用薬 |
JP2003310777A Withdrawn JP2004000281A (ja) | 1992-12-04 | 2003-09-02 | リボザイム増幅診断用薬 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
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JP2003310777A Withdrawn JP2004000281A (ja) | 1992-12-04 | 2003-09-02 | リボザイム増幅診断用薬 |
Country Status (8)
Country | Link |
---|---|
US (1) | US5589332A (ja) |
EP (2) | EP0681613B1 (ja) |
JP (2) | JP3595841B2 (ja) |
AT (1) | ATE217347T1 (ja) |
AU (1) | AU675482B2 (ja) |
DE (1) | DE69331911T2 (ja) |
ES (1) | ES2176233T3 (ja) |
WO (1) | WO1994013833A1 (ja) |
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1993
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- 1993-12-03 WO PCT/US1993/011775 patent/WO1994013833A1/en active IP Right Grant
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EP0681613A4 (en) | 1998-05-20 |
DE69331911D1 (de) | 2002-06-13 |
JP2004000281A (ja) | 2004-01-08 |
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EP1195442A3 (en) | 2002-07-31 |
AU675482B2 (en) | 1997-02-06 |
EP1195442A2 (en) | 2002-04-10 |
DE69331911T2 (de) | 2002-11-21 |
JP3595841B2 (ja) | 2004-12-02 |
ATE217347T1 (de) | 2002-05-15 |
WO1994013833A1 (en) | 1994-06-23 |
AU5739694A (en) | 1994-07-04 |
ES2176233T3 (es) | 2002-12-01 |
EP0681613B1 (en) | 2002-05-08 |
US5589332A (en) | 1996-12-31 |
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