CA2310510C - Procede d'identification et d'inhibition de molecules fonctionnelles d'acide nucleique dans des cellules - Google Patents
Procede d'identification et d'inhibition de molecules fonctionnelles d'acide nucleique dans des cellules Download PDFInfo
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- CA2310510C CA2310510C CA002310510A CA2310510A CA2310510C CA 2310510 C CA2310510 C CA 2310510C CA 002310510 A CA002310510 A CA 002310510A CA 2310510 A CA2310510 A CA 2310510A CA 2310510 C CA2310510 C CA 2310510C
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- 230000035699 permeability Effects 0.000 description 1
- 150000004713 phosphodiesters Chemical class 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 229920000024 polymyxin B Polymers 0.000 description 1
- 229960005266 polymyxin b Drugs 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000006152 selective media Substances 0.000 description 1
- 210000000717 sertoli cell Anatomy 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 238000007860 single-cell PCR Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000002269 spontaneous effect Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 208000006379 syphilis Diseases 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 230000007306 turnover Effects 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
Classifications
-
- 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
-
- 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
- C12Q1/6897—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids involving reporter genes operably linked to promoters
-
- 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
- C12N2310/126—Type of nucleic acid catalytic nucleic acids, e.g. ribozymes involving RNAse P
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Plant Pathology (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
L'invention concerne deux méthodologies. Dans la première, un moyen pour identifier rapidement et efficacement des gènes fonctionnels et essentiels est prévu. Dans la deuxième, un moyen pour produire des molécules nucléiques biologiquement actives (ribozymes, séquences guides externes et antisens) qui peuvent être utilisées pour l'inactivation de gènes fonctionnels. Dans le premier procédé, une banque de séquences guides externes (EGS) à base de toutes les compositions possibles connues est préparée. Dans un mode de réalisation préféré, les EGS constituent douze ou treize mères pour cibler l'RNase bactérien pour le clivage d'un substrat. Cette banque est ajoutée à des cellules contenant les gènes à cribler, par exemple, E. coli. Les cellules dans lesquelles EGS provoque une perte de viabilité, ou d'autre phénotype, sont identifiées. La ou les EGS responsables de la perte de viabilité sont analysées, et l'information relative à la séquence résultante est utilisée pour l'identification du gène dans des séquences génomiques connues. Dans le deuxième procédé, les molécules nucléotidiques ayant une activité biologique optimale, par exemple, dirigeant le clivage d'un gène particulier par RNase P, sont rapidement identifiées au moyen d'un vecteur comprenant deux gènes reporters, le premier en phase avec le gène en question et le deuxième permettant de vérifier que le vecteur est présent dans une cellule ou facilitant la sélection de cellules contenant le vecteurs. Les cellules dans lesquelles le gène en question est clivé par la molécule oligonucléotidique fonctionnelle peuvent ensuite être identifiées en fonction du gène reporter 1. Les molécules oligonucléotidiques fonctionnelles responsables sont ensuite isolées et caractérisées. Lesdits procédés constituent des outils puissants pour l'identification de gènes essentiels dont la séquence est connue seulement comme faisant partie intégrante d'un génome à fonction inconnue, ainsi que des moyens d'identification de molécules oligonucléotidiques fonctionnelles, utiles comme réactifs diagnostiques et comme agents thérapeutiques.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US976,220 | 1997-11-21 | ||
| US08/976,220 US6013447A (en) | 1997-11-21 | 1997-11-21 | Random intracellular method for obtaining optimally active nucleic acid molecules |
| US7985198P | 1998-03-30 | 1998-03-30 | |
| US60/079,851 | 1998-03-30 | ||
| PCT/US1998/024854 WO1999027135A2 (fr) | 1997-11-21 | 1998-11-20 | Procede d'identification et d'inhibition de molecules fonctionnelles d'acide nucleique dans des cellules |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2310510A1 CA2310510A1 (fr) | 1999-06-03 |
| CA2310510C true CA2310510C (fr) | 2007-04-17 |
Family
ID=26762503
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002310510A Expired - Fee Related CA2310510C (fr) | 1997-11-21 | 1998-11-20 | Procede d'identification et d'inhibition de molecules fonctionnelles d'acide nucleique dans des cellules |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP1032707A2 (fr) |
| JP (1) | JP2001524317A (fr) |
| AU (1) | AU732321B2 (fr) |
| CA (1) | CA2310510C (fr) |
| WO (1) | WO1999027135A2 (fr) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1034308A4 (fr) * | 1997-12-04 | 2002-04-03 | Smithkline Beecham Corp | Procede mettant en oeuvre des sequences antisens d'expression pour produire des cellules mutantes exprimees de maniere conditionnelle |
| AU4186600A (en) * | 1999-03-31 | 2000-10-16 | Rosetta Inpharmatics, Inc. | Essential genes of yeast as targets for antifungal agents, herbicides, insecticides and anti-proliferation drugs |
| GB9912965D0 (en) * | 1999-06-03 | 1999-08-04 | Oxford Biomedica Ltd | In vivo selection method |
| GB9929744D0 (en) * | 1999-12-17 | 2000-02-09 | Univ Nottingham | Modifying micro-organisms |
| WO2001048239A2 (fr) * | 1999-12-23 | 2001-07-05 | Xantos Biomedicine Ag | Procede d'echantillonnage pour acides nucleiques |
| AU2008202554C1 (en) * | 2001-02-23 | 2011-07-07 | Dsm Ip Assets B.V. | Novel genes encoding novel proteolytic enzymes |
| US20130244905A1 (en) * | 2010-07-06 | 2013-09-19 | Ed Grabczyk | Reporter for RNA Polymerase II Termination |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5168053A (en) * | 1989-03-24 | 1992-12-01 | Yale University | Cleavage of targeted RNA by RNAase P |
| EP0638121B1 (fr) * | 1992-04-28 | 1996-07-17 | Yale University | Clivage cible d'arn au moyen de ribonuclease eucaryote p et d'une sequence-guide externe |
| JPH08507203A (ja) * | 1992-12-04 | 1996-08-06 | イノーバー ラボラトリーズ，インコーポレイテッド | 調節可能な核酸治療およびそれらの使用方法 |
| WO1996018733A2 (fr) * | 1994-12-14 | 1996-06-20 | Innovir Laboratories, Inc. | Inactivation induite par ribozyme de l'arn associe a la leucemie |
| WO1997010360A1 (fr) * | 1995-09-13 | 1997-03-20 | Chiron Corporation | Procede et produit d'assemblage pour la recherche d'inhibiteurs d'activation de la transcription |
| US5877162A (en) * | 1996-03-14 | 1999-03-02 | Innovir Laboratories, Inc. | Short external guide sequences |
| WO1998006837A1 (fr) * | 1996-08-16 | 1998-02-19 | Yale University | Transformation du phenotype de bacteries chimio-resistantes en phenotype chimio-sensible |
| AU5927598A (en) * | 1997-01-23 | 1998-08-18 | Immusol Incorporated | Gene functional analysis and discovery using randomized or target-specific ribozyme gene vector libraries |
-
1998
- 1998-11-20 EP EP98959542A patent/EP1032707A2/fr not_active Withdrawn
- 1998-11-20 CA CA002310510A patent/CA2310510C/fr not_active Expired - Fee Related
- 1998-11-20 AU AU15323/99A patent/AU732321B2/en not_active Ceased
- 1998-11-20 JP JP2000522276A patent/JP2001524317A/ja active Pending
- 1998-11-20 WO PCT/US1998/024854 patent/WO1999027135A2/fr not_active Ceased
Also Published As
| Publication number | Publication date |
|---|---|
| AU732321B2 (en) | 2001-04-12 |
| AU1532399A (en) | 1999-06-15 |
| WO1999027135A2 (fr) | 1999-06-03 |
| JP2001524317A (ja) | 2001-12-04 |
| CA2310510A1 (fr) | 1999-06-03 |
| EP1032707A2 (fr) | 2000-09-06 |
| WO1999027135A3 (fr) | 1999-07-29 |
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| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20141120 |