WO2012110899A3 - Method for localizedin situdetection of mrna - Google Patents

Method for localizedin situdetection of mrna Download PDF

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Publication number
WO2012110899A3
WO2012110899A3 PCT/IB2012/000995 IB2012000995W WO2012110899A3 WO 2012110899 A3 WO2012110899 A3 WO 2012110899A3 IB 2012000995 W IB2012000995 W IB 2012000995W WO 2012110899 A3 WO2012110899 A3 WO 2012110899A3
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WO
WIPO (PCT)
Prior art keywords
rna
padlock probe
cdna
situ
situdetection
Prior art date
Application number
PCT/IB2012/000995
Other languages
French (fr)
Other versions
WO2012110899A2 (en
Inventor
Mats Nilsson BERNITZ
Chatarina LARSSON
Ida GRUNDBERG
Original Assignee
Bernitz Mats Nilsson
Larsson Chatarina
Grundberg Ida
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Bernitz Mats Nilsson, Larsson Chatarina, Grundberg Ida filed Critical Bernitz Mats Nilsson
Priority to CN201280009026.1A priority Critical patent/CN103649331A/en
Priority to EP12728767.0A priority patent/EP2675916B1/en
Priority to BR112013020773-6A priority patent/BR112013020773B1/en
Priority to JP2013554022A priority patent/JP5881746B2/en
Priority to AU2012219132A priority patent/AU2012219132B2/en
Priority to CA2827497A priority patent/CA2827497C/en
Publication of WO2012110899A2 publication Critical patent/WO2012110899A2/en
Publication of WO2012110899A3 publication Critical patent/WO2012110899A3/en

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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
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • 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
    • C12Q1/6809—Methods for determination or identification of nucleic acids involving differential detection
    • 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
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • 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
    • C12Q1/6841—In situ hybridisation
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/112—Disease subtyping, staging or classification
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/16—Primer sets for multiplex assays

Abstract

The present invention relates to the detection of RNA in a sample of cells. More particularly, the present invention relates to the localized detection of RNA in situ. The method relies on the conversion of RNA to complementary DNA prior to the targeting of the cDNA with a padlock probe(s). The hybridization of the padlock probe(s) relies on the nucleotide sequence of the cDNA which is derived from the corresponding nucleotide sequence of the target RNA. Rolling circle amplification of the subsequently circularized padlock probe produces a rolling circle product which may be detected. Advantageously, this allows the RNA to be detected in situ.
PCT/IB2012/000995 2011-02-15 2012-02-15 Method for localizedin situdetection of mrna WO2012110899A2 (en)

Priority Applications (6)

Application Number Priority Date Filing Date Title
CN201280009026.1A CN103649331A (en) 2011-02-15 2012-02-15 Method for localizedin situdetection of MRNA
EP12728767.0A EP2675916B1 (en) 2011-02-15 2012-02-15 METHOD FOR LOCALIZED IN SITU DETECTION OF mRNA
BR112013020773-6A BR112013020773B1 (en) 2011-02-15 2012-02-15 METHOD FOR LOCALIZED IN SITU DETECTION OF RNA, METHOD FOR DETERMINING THE PRESENCE AND LOCATION OF A GENETIC SEQUENCE AND METHOD FOR IN SITU LOCATION AND/OR DETECTION OF A SPECIFIC RNA
JP2013554022A JP5881746B2 (en) 2011-02-15 2012-02-15 A method for localized in situ detection of mRNA
AU2012219132A AU2012219132B2 (en) 2011-02-15 2012-02-15 Method for localized in situ detection of mRNA
CA2827497A CA2827497C (en) 2011-02-15 2012-02-15 Method for localized in situ detection of mrna

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US201161442921P 2011-02-15 2011-02-15
US61/442,921 2011-02-15
US201161473662P 2011-04-08 2011-04-08
US61/473,662 2011-04-08

Publications (2)

Publication Number Publication Date
WO2012110899A2 WO2012110899A2 (en) 2012-08-23
WO2012110899A3 true WO2012110899A3 (en) 2012-12-13

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PCT/IB2012/000995 WO2012110899A2 (en) 2011-02-15 2012-02-15 Method for localizedin situdetection of mrna

Country Status (8)

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US (2) US8551708B2 (en)
EP (1) EP2675916B1 (en)
JP (2) JP5881746B2 (en)
CN (2) CN113151404A (en)
AU (1) AU2012219132B2 (en)
BR (1) BR112013020773B1 (en)
CA (1) CA2827497C (en)
WO (1) WO2012110899A2 (en)

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