FR2997705B1 - NUCLEOTIDE 3 'BLOCKING TRANSLATION OF PROTEINS (SASB) - Google Patents
NUCLEOTIDE 3 'BLOCKING TRANSLATION OF PROTEINS (SASB)Info
- Publication number
- FR2997705B1 FR2997705B1 FR1202997A FR1202997A FR2997705B1 FR 2997705 B1 FR2997705 B1 FR 2997705B1 FR 1202997 A FR1202997 A FR 1202997A FR 1202997 A FR1202997 A FR 1202997A FR 2997705 B1 FR2997705 B1 FR 2997705B1
- Authority
- FR
- France
- Prior art keywords
- blocking
- sasb
- nucleic acid
- synthesis
- peptide
- 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.)
- Active
Links
- 230000000903 blocking effect Effects 0.000 title abstract 10
- 108090000623 proteins and genes Proteins 0.000 title abstract 5
- 239000002773 nucleotide Substances 0.000 title abstract 3
- 102000004169 proteins and genes Human genes 0.000 title abstract 3
- 108020004999 messenger RNA Proteins 0.000 abstract 6
- 108020004707 nucleic acids Proteins 0.000 abstract 6
- 102000039446 nucleic acids Human genes 0.000 abstract 6
- 150000007523 nucleic acids Chemical class 0.000 abstract 5
- 108090000765 processed proteins & peptides Proteins 0.000 abstract 4
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 abstract 4
- 108020004705 Codon Proteins 0.000 abstract 3
- 230000015572 biosynthetic process Effects 0.000 abstract 3
- 230000000295 complement effect Effects 0.000 abstract 3
- 238000003786 synthesis reaction Methods 0.000 abstract 3
- 230000014616 translation Effects 0.000 abstract 3
- 108020005038 Terminator Codon Proteins 0.000 abstract 2
- 238000000034 method Methods 0.000 abstract 2
- 125000003729 nucleotide group Chemical group 0.000 abstract 2
- 238000001243 protein synthesis Methods 0.000 abstract 2
- 229950010131 puromycin Drugs 0.000 abstract 2
- -1 N-3 nucleic acids Chemical class 0.000 abstract 1
- 230000000692 anti-sense effect Effects 0.000 abstract 1
- 210000004899 c-terminal region Anatomy 0.000 abstract 1
- 238000003384 imaging method Methods 0.000 abstract 1
- 230000005764 inhibitory process Effects 0.000 abstract 1
- 230000014759 maintenance of location Effects 0.000 abstract 1
- 210000003705 ribosome Anatomy 0.000 abstract 1
- 230000008685 targeting Effects 0.000 abstract 1
- 230000001988 toxicity Effects 0.000 abstract 1
- 231100000419 toxicity Toxicity 0.000 abstract 1
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
-
- 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
- C12N15/62—DNA sequences coding for fusion proteins
-
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
-
- 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
- C12N2310/11—Antisense
-
- 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
Abstract
Cette invention décrit un procédé permettant de bloquer réversiblement la synthèse d'une protéine cible dans un système eucaryote ou procaryote en ciblant l'ARNm de du gène correspondant. Elle permet également de bloquer réversiblement la synthèse de toutes les protéines dans un système en bloquant au moins un gène spécifiquement exprimé dans ledit système sous sa forme d'ARN messager. Le procédé consiste en au moins un acide nucléique dit bloquant (1), ou séquence anti-sens bloquant (SASB) ou une combinaison quelconque de SASB, un SASB donné présentant d'une taille N comprise entre 3 à 50 nucléotides, tel que chaque acide nucléique bloquant présente à l'extrémité 3', trois nucléotides formant un anti-codon de terminaison (2) de la traduction (« anti-ter ») de séquence complémentaire à un codon de terminaison de traduction (3) («codon ter ») et plus particulièrement complémentaire au codon ter de son ARNm cible, les N-3 nucléotides restant (5), dudit acide nucléique bloquant, étant complémentaires des N-3 acides nucléiques suivant le codon stop de l'ARNm cible. Un Acide nucléique bloquant, en s'hybridant sur son ARNm cible, va bloquer l'action des facteurs de relargage du ribosome, interrompant la synthèse protéique en séquestrant le ribosome dans un complexe ARNm/acide nucléique bloquant/peptide en cours de synthèse. Lorsque suffisamment de SASB sont présents dans le système, le blocage de tous les ribosomes conduit à l'arrêt de la synthèse protéique. Le ribosome et le peptide bloqués pourront être relargués par l'action de la puromycine. La puromycine, étant intégrée en C terminal du peptide au moment du relargage permet d'intégrer au peptide un insert présentant différents types de fonctions et notamment des fonctions de toxicité, de contraste pour l'imagerie ou d'inhibition.This invention describes a method for reversibly blocking the synthesis of a target protein in a eukaryotic or prokaryotic system by targeting the mRNA of the corresponding gene. It also makes it possible to reversibly block the synthesis of all the proteins in a system by blocking at least one gene specifically expressed in said system in its form of messenger RNA. The method consists of at least one so-called blocking nucleic acid (1), or antisense blocking sequence (SASB) or any combination of SASB, a given SASB having a size N of between 3 to 50 nucleotides, such that each blocking nucleic acid present at the 3 'end, three nucleotides forming an anti-translation termination codon (2) ("anti-ter") with a sequence complementary to a translation termination codon (3) ("codon ter ") And more particularly complementary to the ter codon of its target mRNA, the remaining N-3 nucleotides (5), of said blocking nucleic acid, being complementary to the N-3 nucleic acids following the stop codon of the target mRNA. A blocking nucleic acid, by hybridizing to its target mRNA, will block the action of ribosome release factors, interrupting protein synthesis by sequestering the ribosome in an mRNA / blocking nucleic acid / peptide complex during synthesis. When enough SASB is present in the system, blocking all ribosomes leads to the cessation of protein synthesis. The blocked ribosome and peptide can be released by the action of puromycin. The puromycin, being integrated at the C terminal of the peptide at the time of release, makes it possible to integrate into the peptide an insert having different types of functions and in particular functions of toxicity, contrast for imaging or inhibition.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR1202997A FR2997705B1 (en) | 2012-11-08 | 2012-11-08 | NUCLEOTIDE 3 'BLOCKING TRANSLATION OF PROTEINS (SASB) |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR1202997A FR2997705B1 (en) | 2012-11-08 | 2012-11-08 | NUCLEOTIDE 3 'BLOCKING TRANSLATION OF PROTEINS (SASB) |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| FR2997705A1 FR2997705A1 (en) | 2014-05-09 |
| FR2997705B1 true FR2997705B1 (en) | 2015-10-16 |
Family
ID=48771472
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| FR1202997A Active FR2997705B1 (en) | 2012-11-08 | 2012-11-08 | NUCLEOTIDE 3 'BLOCKING TRANSLATION OF PROTEINS (SASB) |
Country Status (1)
| Country | Link |
|---|---|
| FR (1) | FR2997705B1 (en) |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030096775A1 (en) * | 2001-10-23 | 2003-05-22 | Isis Pharmaceuticals Inc. | Antisense modulation of complement component C3 expression |
| WO2006017522A2 (en) * | 2004-08-03 | 2006-02-16 | University Of Utah Research Foundation | Use of antisense oligonucleotides to effect translation modulation |
| WO2006082058A2 (en) * | 2005-02-02 | 2006-08-10 | Universität Bayreuth | Cell-free translation system for the production of (chemically) modified proteins |
| US20110171287A1 (en) * | 2007-11-05 | 2011-07-14 | Baltic Technology Develpment, Ltd. | Use of Oligonucleotides with Modified Bases as Antiviral Agents |
| CN102197136B (en) * | 2008-08-28 | 2016-09-28 | 诺华股份有限公司 | Cell surface display by the polypeptide isoforms of termination codon liaison |
-
2012
- 2012-11-08 FR FR1202997A patent/FR2997705B1/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| FR2997705A1 (en) | 2014-05-09 |
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