DK2225393T3 - Fremgangsmåde til hybridisering af nukleinsyrer - Google Patents

Fremgangsmåde til hybridisering af nukleinsyrer Download PDF

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Publication number
DK2225393T3
DK2225393T3 DK08868164.8T DK08868164T DK2225393T3 DK 2225393 T3 DK2225393 T3 DK 2225393T3 DK 08868164 T DK08868164 T DK 08868164T DK 2225393 T3 DK2225393 T3 DK 2225393T3
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Denmark
Prior art keywords
oligonucleotide
probe
conjugate
oligocation
nucleic acid
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DK08868164.8T
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English (en)
Inventor
Patrick Erbacher
Nathalie Lenne
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Polyplus Transfection
Univ Strasbourg
Centre Nat Rech Scient
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Publication of DK2225393T3 publication Critical patent/DK2225393T3/da

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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
    • C12Q1/6832—Enhancement of hybridisation reaction
    • 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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • 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

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Genetics & Genomics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biomedical Technology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Claims (13)

1. Fremgangsmåde til detektering af en target-nukleinsyre i en prøve ved hybridisering med et oligonukleotid-oligokationskonjugat, hvilken fremgangsmåde omfatter det at gøre det muligt for nukleinsyren at reagere med et oligonukleotid-oligokationskonjugat omfattende mindst Ai- og Bj-enheder, som er forbundet med hinanden direkte eller via en linker-gruppe, hvor . A, er et i-mer-oligonukleotid med i = 3 til 50, hvor A, er en oligomer med naturligt eller ikke naturligt forekommende nukleobaser og/eller pentafuranosyl-grupper og/eller native phosphodiesterbindinger, eventuelt omfattende en markørgruppe, . Bj er en j-mer organisk oligokationsenhed med j = 1 til 50, hvor B er HP03-Ri-(NH-R2)n-NH-R3-0-, hvor R1; R2 og R3 er identiske eller forskellige, er et eventuelt substitueret C1-C6 lineært, forgrenet eller cyklisk alkylenradi-kal, hvilke NH-R2-enheder er identiske eller forskellige, når n er >1; eller HP03-Ri-CH(X)-R3-0-, hvor R-\ og R3 er identiske eller forskellige, er et eventuelt substitueret C1-C6 lineært, forgrenet eller cyklisk alkylenradikal, og X er en putrescin-, spermidin- eller sperminrest, hvor strukturen af konjugatet er struktur IV hvor
- R4 og R6 er identiske eller forskellige, er H eller en linker, en quencher, en markør, en chromophor- eller fluorophor-gruppe, biotin, hydrofob kæde, kolesterolderivat, antigen, protein, peptid, sukker eller phosphatgruppe; hvor target-nukleinsyren er en specifik sekvens i et helt genom; og hvor fremgangsmåden omfatter - anvendelse af konjugatet som en hybridisering eller dobbeltmarkeret sonde i et realtids-PCR-assay omfattende en DNA- eller RNA-polymerase i nærvær af hele genomet.
2. Fremgangsmåde ifølge krav 1, hvorved hele genomet er genomisk DNA.
3. Fremgangsmåde ifølge krav 1 eller 2, hvorved nukleotiderne af A, udvælges fra gruppen bestående af deoxyribonukleotider, ribonukleotider, locked (LNA) nukleotider, PNA samt deres kemiske modificeringer eller substitutioner ved phosphorothioat, 2'-fluor- eller 2'-0-alkylgrupper.
4. Fremgangsmåde ifølge et af kravene 1-3, hvorved konjugatet anvendes som en dobbeltmarkeret sonde i realtids-PCR-assayet.
5. Fremgangsmåde ifølge et af kravene 1-4, hvorved B er ΡΙΡ03-(ΟΡΙ2)4- NH2+-(CH2)3-NH2+-(CH2)4-NH2+-(CH2)3-NH2+-(CH2)4-0-.
6. Fremgangsmåde ifølge krav 5, hvor j = 4.
7. Fremgangsmåde ifølge krav 5 eller 6, hvorved R4 er en quencher, og R6 er en fluorophor.
8. Fremgangsmåde ifølge et af kravene 5-7, hvor nukleotiderne af A, er deoxyribonukleotider.
9. Fremgangsmåde ifølge et af kravene 1-8, hvor mindst et oligonukleotid-oligokationskonjugat anvendes som en sonde til at skelne mellem en vildtype- og en mutant target-nukleinsyre.
10. Fremgangsmåde ifølge et af kravene 1-8, hvor mindst et oligonukleotid-oligokationskonjugat anvendes som en sonde til allel diskrimination.
11. Fremgangsmåde ifølge et af kravene 1-8, hvor mindst et oligonukleotid-oligokationskonjugat anvendes som en sonde til detektering af mutationer.
12. Fremgangsmåde ifølge et af kravene 1-8 og 11, hvor mindst et oligonukleotid-oligokationskonjugat anvendes som en sonde til detektering af SNP.
13. Fremgangsmåde ifølge et af kravene 1-12, hvor konjugatet anvendes som en realtids-detekteringssonde i et 5' nuklease-assay designet til at am-plificere det humane faktor V-gen.
DK08868164.8T 2007-12-27 2008-09-12 Fremgangsmåde til hybridisering af nukleinsyrer DK2225393T3 (da)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US914507P 2007-12-27 2007-12-27
EP07025148A EP2075342A1 (en) 2007-12-27 2007-12-27 Method for hybridizing nucleic acids
PCT/IB2008/002789 WO2009083763A1 (en) 2007-12-27 2008-09-12 Method for hybridizing nucleic acids

Publications (1)

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DK2225393T3 true DK2225393T3 (da) 2015-05-26

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US (1) US8465920B2 (da)
EP (2) EP2075342A1 (da)
JP (1) JP2011507542A (da)
KR (1) KR20100124705A (da)
CN (1) CN101978071A (da)
AU (1) AU2008345460A1 (da)
BR (1) BRPI0821698A2 (da)
CA (1) CA2709183A1 (da)
DK (1) DK2225393T3 (da)
ES (1) ES2537625T3 (da)
IL (1) IL206542A0 (da)
NZ (1) NZ586373A (da)
RU (1) RU2010131166A (da)
WO (1) WO2009083763A1 (da)

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EP2542678B1 (en) 2010-03-04 2017-04-12 InteRNA Technologies B.V. A MiRNA MOLECULE DEFINED BY ITS SOURCE AND ITS THERAPEUTIC USES IN CANCER ASSOCIATED WITH EMT
EP2591106A1 (en) 2010-07-06 2013-05-15 InteRNA Technologies B.V. Mirna and its diagnostic and therapeutic uses in diseases or conditions associated with melanoma, or in diseases or conditions associated with activated braf pathway
WO2012033190A1 (ja) * 2010-09-10 2012-03-15 三菱化学メディエンス株式会社 光を用いた核酸の増幅抑制方法および高感度な選択的核酸増幅方法
EP2474617A1 (en) 2011-01-11 2012-07-11 InteRNA Technologies BV Mir for treating neo-angiogenesis
EP3369818B1 (en) 2011-12-22 2021-06-09 InteRNA Technologies B.V. Mirna for treating head and neck cancer
KR102146523B1 (ko) * 2012-01-06 2020-08-20 조원창 표적 핵산의 향상된 증폭
EP2917348A1 (en) 2012-11-06 2015-09-16 InteRNA Technologies B.V. Combination for use in treating diseases or conditions associated with melanoma, or treating diseases or conditions associated with activated b-raf pathway
US9410172B2 (en) * 2013-09-16 2016-08-09 General Electric Company Isothermal amplification using oligocation-conjugated primer sequences
DE102017123919A1 (de) 2017-10-13 2019-04-18 Gna Biosolutions Gmbh Verfahren und Vorrichtung zur Lyse von Mikroorganismen
CN111566212A (zh) 2017-11-03 2020-08-21 因特尔纳技术有限公司 miRNA分子，等同物，安塔够妙或其来源用于治疗和/或诊断与神经元缺陷相关的病症和/或疾病或用于神经元生成和/或再生
JP2021513508A (ja) 2018-02-12 2021-05-27 インテアールエヌエー テクノロジーズ ビー．ヴイ．ＩｎｔｅＲＮＡ Ｔｅｃｈｎｏｌｏｇｉｅｓ Ｂ．Ｖ． 抗がんマイクロｒｎａ及びその脂質製剤
KR20200045212A (ko) 2018-10-22 2020-05-04 (주)바이오니아 옥타민 또는 옥타민 유도체가 결합된 프로브 및 이의 용도
CN113444778A (zh) 2020-03-27 2021-09-28 广州达安基因股份有限公司 具有高杂交性能的寡核苷酸缀合物及其应用
IT202200026595A1 (it) 2022-12-22 2024-06-22 Fond Telethon Ets Nuovi inibitori di regolatori epigenetici

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EP1013770A1 (en) * 1998-12-23 2000-06-28 Université Louis Pasteur de Strasbourg Non-viral transfection vector
WO2006052854A2 (en) * 2004-11-04 2006-05-18 Raul Andino Syntheses of polyamine conjugates of small interfering rnas (si-rnas) and conjugates formed thereby
NZ569138A (en) 2005-12-15 2012-06-29 Centre Nat Rech Scient Cationic oligonucleotides automated methods for preparing same and their uses

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RU2010131166A (ru) 2012-02-10
EP2075342A1 (en) 2009-07-01
EP2225393A1 (en) 2010-09-08
KR20100124705A (ko) 2010-11-29
CN101978071A (zh) 2011-02-16
NZ586373A (en) 2012-09-28
WO2009083763A1 (en) 2009-07-09
AU2008345460A1 (en) 2009-07-09
JP2011507542A (ja) 2011-03-10
CA2709183A1 (en) 2009-07-09
US8465920B2 (en) 2013-06-18
BRPI0821698A2 (pt) 2015-06-16
US20100311056A1 (en) 2010-12-09
IL206542A0 (en) 2010-12-30
ES2537625T3 (es) 2015-06-10
EP2225393B1 (en) 2015-02-25

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