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BR9914773A - Set of generic elements, method for the production of single-stranded DNA, mrna transcription, nucleic acid construction, ssdna transcription, vector, vector system, host cell, set for the production of a single-stranded nucleic acid sequence, method for in vivo or in vitro production of a single-stranded nucleic acid sequence, transcription of single-stranded cdna, inhibitory nucleic acid, heteroduplex molecule, and pharmaceutical composition - Google Patents

Set of generic elements, method for the production of single-stranded DNA, mrna transcription, nucleic acid construction, ssdna transcription, vector, vector system, host cell, set for the production of a single-stranded nucleic acid sequence, method for in vivo or in vitro production of a single-stranded nucleic acid sequence, transcription of single-stranded cdna, inhibitory nucleic acid, heteroduplex molecule, and pharmaceutical composition

Info

Publication number
BR9914773A
BR9914773A BR9914773-4A BR9914773A BR9914773A BR 9914773 A BR9914773 A BR 9914773A BR 9914773 A BR9914773 A BR 9914773A BR 9914773 A BR9914773 A BR 9914773A
Authority
BR
Brazil
Prior art keywords
nucleic acid
stranded
cdna
transcription
production
Prior art date
Application number
BR9914773-4A
Other languages
Portuguese (pt)
Inventor
Charles A Conrad
Original Assignee
Ingene Inc
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 Ingene Inc filed Critical Ingene Inc
Publication of BR9914773A publication Critical patent/BR9914773A/en

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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113Non-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
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/10Type of nucleic acid
    • C12N2310/11Antisense
    • C12N2310/111Antisense spanning the whole gene, or a large part of it
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/10Type of nucleic acid
    • C12N2310/12Type of nucleic acid catalytic nucleic acids, e.g. ribozymes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/50Physical structure
    • C12N2310/53Physical structure partially self-complementary or closed

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

"CONJUNTO DE ELEMENTOS GENéTICOS, MéTODO PARA A PRODUçãO DE DNA DE CORDãO úNICO, TRANSCRIçãO DE mRNA, CONSTRUçãO DE áCIDO NUCLéICO, TRANSCRIçãO DE ssDNA, VETOR, SISTEMA VETOR, CéLULA HOSPEDEIRA, CONJUNTO PARA A PRODUçãO DE UMA SEQuêNCIA DE áCIDO NUCLéICO DE CORDãO úNICO, MéTODO PARA A PRODUçãO IN VIVO OU IN VITRO DE UMA SEQuêNCIA DE áCIDO NUCLéICO DE CORDãO úNICO, TRANSCRIçãO DE CDNA DE CORDãO úNICO, áCIDO NUCLéICO INIBIDOR, MOLéCULA HETERODUPLEX E COMPOSIçãO FARMACêUTICA". Métodos e composições para a produção de cDNA de um único filamento (ss-cDNA) em células eucarióticas, especificamente um cassete de DNA que produz s-cDNA in vivo O cassete contém o gene de codificação de transcriptase/Rnase H reversa do vírus de leucemia de murinos de Moloney, um gene de endonuclease de restrição de bactérias, e uma seq³ência de interesse que produz um modelo de RNA a partir do qual a transcriptase reversa sintetiza o ss-cDNA de uma seq³ência específica. O ss-cDNA é então modificado para remover todas as seq³ências de vetores de flanqueamento ao tirar vantagem da estrutura de "haste-laço" do ss-cDNA, que se forma como resultado da inclusão de uma repetição em tandem invertida que permite que o ss-cDNA de dobre de volta sobre si mesmo, formando uma haste de DNA de filamento duplo, na seq³ência de interesse. A haste de filamento duplo contém um ou mais elementos genéticos funcionais, tais como um sítio de reconhecimento de endonuclease de restrição e o laço, que continua como ss-cDNA, compreende qualquer seq³ência de nucleotídeos desejada. Esse desenho permite que a haste de filamento duplo do intermediário de haste-laço a ser clivada pela(s) endonuclease(s) de restrição correspondente(s) desejada(s) específica(s) para o sítio na haste e na porção do laço, ou seq³ência de interesse, seja então liberada como um pedaço de DNA de um único filamento linearizado. Esse pedaço de ss-DNA liberado (ou clivado) contém informações mínimas, caso haja alguma, sobre a seq³ência tanto a montante da posição 51 quanto a jusante da posição 3' em relação à porção da haste de filamento duplo anterior que contém o sítio de corte de endonuclease de restrição. As transfecções in vivo utilizando o sistema de vetores de DNA aqui descrito demonstram o uso desse sistema para produzir ss-DNA em células hospedeiras."SET OF GENETIC ELEMENTS, METHOD FOR THE PRODUCTION OF SINGLE CORD DNA, MRNA TRANSCRIPTION, NUCLEIC ACID CONSTRUCTION, SDSNA TRANSCRIPTION, VECTOR, VECTOR SYSTEM, HOSPITAL CELL, NUCLEUM CUTTING UNIT , METHOD FOR THE IN VIVO OR IN VITRO PRODUCTION OF A SINGLE CORDANIC NUCLEIC ACID, TRANSCRIPTION OF SINGLE CORDANIC CDNA, INHIBITOR NUCLEIC ACID, HETERODUPLEX MOLECULE AND PHARMACEUTICAL COMPOSITION ". Methods and compositions for the production of single-strand cDNA (ss-cDNA) in eukaryotic cells, specifically a DNA cassette that produces s-cDNA in vivo The cassette contains the leukemia virus reverse transcriptase / R Hase encoding gene of Moloney murines, a bacterial restriction endonuclease gene, and a sequence of interest that produces an RNA model from which reverse transcriptase synthesizes the ss-cDNA of a specific sequence. The ss-cDNA is then modified to remove all flanking vector sequences by taking advantage of the ss-cDNA "rod-loop" structure, which is formed as a result of including an inverted tandem repeat that allows the ss -cDNA fold back on itself, forming a double-stranded DNA strand, in the sequence of interest. The double stranded rod contains one or more functional genetic elements, such as a restriction endonuclease recognition site and the loop, which continues as ss-cDNA, comprises any desired nucleotide sequence. This design allows the double-stranded rod of the rod-loop intermediate to be cleaved by the desired corresponding restriction endonuclease (s) specific to the site on the rod and portion of the loop , or sequence of interest, is then released as a piece of DNA from a single linearized strand. This piece of ss-DNA released (or cleaved) contains minimal information, if any, about the sequence both upstream of position 51 and downstream of position 3 'in relation to the portion of the anterior double-stranded rod containing the site of restriction endonuclease cut. In vivo transfections using the DNA vector system described here demonstrate the use of this system to produce ss-DNA in host cells.

BR9914773-4A 1998-10-09 1999-10-12 Set of generic elements, method for the production of single-stranded DNA, mrna transcription, nucleic acid construction, ssdna transcription, vector, vector system, host cell, set for the production of a single-stranded nucleic acid sequence, method for in vivo or in vitro production of a single-stranded nucleic acid sequence, transcription of single-stranded cdna, inhibitory nucleic acid, heteroduplex molecule, and pharmaceutical composition BR9914773A (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US16979398A 1998-10-09 1998-10-09
US39778299A 1999-09-16 1999-09-16
PCT/US1999/023936 WO2000022114A1 (en) 1998-10-09 1999-10-12 PRODUCTION OF ssDNA $i(IN VIVO)

Publications (1)

Publication Number Publication Date
BR9914773A true BR9914773A (en) 2002-02-05

Family

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Family Applications (1)

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BR9914773-4A BR9914773A (en) 1998-10-09 1999-10-12 Set of generic elements, method for the production of single-stranded DNA, mrna transcription, nucleic acid construction, ssdna transcription, vector, vector system, host cell, set for the production of a single-stranded nucleic acid sequence, method for in vivo or in vitro production of a single-stranded nucleic acid sequence, transcription of single-stranded cdna, inhibitory nucleic acid, heteroduplex molecule, and pharmaceutical composition

Country Status (5)

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AU (1) AU6298899A (en)
BR (1) BR9914773A (en)
CA (1) CA2345936A1 (en)
MX (1) MXPA01003643A (en)
WO (1) WO2000022114A1 (en)

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