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WO2019000148A1 - Arnsi du gène abcb6 humain et utilisation correspondante - Google Patents

Arnsi du gène abcb6 humain et utilisation correspondante Download PDF

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Publication number
WO2019000148A1
WO2019000148A1 PCT/CN2017/089928 CN2017089928W WO2019000148A1 WO 2019000148 A1 WO2019000148 A1 WO 2019000148A1 CN 2017089928 W CN2017089928 W CN 2017089928W WO 2019000148 A1 WO2019000148 A1 WO 2019000148A1
Authority
WO
WIPO (PCT)
Prior art keywords
sirna
abcb6 gene
gene
abcb6
sequence
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.)
Ceased
Application number
PCT/CN2017/089928
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English (en)
Chinese (zh)
Inventor
毛吉炎
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shenzhen Biocan Technologies Co Ltd
Original Assignee
Shenzhen Biocan Technologies Co Ltd
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 Shenzhen Biocan Technologies Co Ltd filed Critical Shenzhen Biocan Technologies Co Ltd
Priority to PCT/CN2017/089928 priority Critical patent/WO2019000148A1/fr
Publication of WO2019000148A1 publication Critical patent/WO2019000148A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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

Definitions

  • the present invention belongs to the field of molecular genetics, and particularly relates to an siRNA capable of inhibiting the expression of a human ABCB6 gene and an application thereof.
  • Adenosine triphosphate binding cassette transporter is a large class of transmembrane proteins that utilize the energy of hydrolyzing A TP to transport transmembrane transport of various endogenous and exogenous biomolecules in solute. Substrates include: sugars, amino acids, metal ions, peptides, proteins, cellular metabolites, and drugs. ABCB transporters are widely present in eukaryotic and prokaryotic organisms. To date, 49 ABC transgenic superfamily members have been identified in the human genome, which are divided into seven subfamilies of A-G.
  • ABCB1 is the first human ABC transporter to be discovered. Because multiple members are associated with multidrug resistance (MD R), the ABCB subfamily is also known as the ABC transporter MDR family. As one of them, A BCB6 is not only related to MDR, but also related to the pathogenesis of various tumors. However, the function and role of ABCB6 in it have not been clarified so far, and further research is needed.
  • RNA interference RNA interference
  • RNAi small interfering RNA
  • siABCB6 sequence is as follows: [0007] Justice Chain: 5'- GGCAUCUGGAUCAAGUUCA -3' (SEQ ID NO: 1)
  • Antisense strand 5,- UGAACUUGAUCCAGAUGCC -3, (SEQ ID NO: 2).
  • the present invention provides siABCB6 which has the advantages of high interference efficiency, high-efficiency and specific inhibition of ABCB6 gene expression, and can be used as a powerful tool for the preparation of a medicament for treating a disease associated with abnormal expression of ABCB6 gene.
  • 1 is a schematic diagram showing the results of quantitative PCR detection of ABCB6 gene expression levels after transfection of A375 cells with siABCB6.
  • A375 cells purchased from Biovector Plasmid Vector Culture Cell Gene Collection
  • DMEM complete medium containing 10% fetal bovine serum
  • 6-well plates were plated at a ratio of 150,000 cells/well at 37°C. C, 5 % C02 was cultured for 18 h.
  • Cell transfection was performed using the Lipofectamine 3000 Transfection Kit (Invitrogen) according to the product instructions.
  • RNA extraction Total RNA from normal and transfected siABCB6 ABCB6 cells was extracted using the QIAGEN RNeasy Mini Kit.
  • Reverse Transcription Reverse transcription was performed using FastQuant RT Super Mix.
  • Quantitative PCR was carried out, and the reaction system was 20 ⁇ , and 1 L cDNA was added as a template for each reaction.
  • the reaction procedure is: (1) 95 °C 30 s, (2) 95 °C 5s, (3) 60 °C 30s, (2)-(3), 40 cycles. With GAPDH as the internal reference, the results are shown in Figure 1.
  • the quantitative PCR primers used are shown in Table 1:
  • the mRNA expression level of ABCB6 gene in A375 cells transfected with siABCB6 was significantly decreased compared with normal A375 cells, indicating that siABCB6 of the present invention can efficiently and specifically inhibit the expression of ABCB6 gene.
  • the siABCB6 provided by the invention has high interference efficiency and can efficiently and specifically inhibit the ABCB6 gene expression.
  • the advantages of the drug can be used as a powerful tool for the preparation of drugs for the treatment of diseases related to abnormal expression of ABCB6 gene.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Public Health (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne un fragment d'ARN interférant qui inhibe efficacement et spécifiquement le niveau d'expression d'ARNm du gène ABCB6, ledit fragment pouvant être utilisé pour préparer un médicament pour traiter une maladie associée à l'expression anormale du gène ABCB6.
PCT/CN2017/089928 2017-06-26 2017-06-26 Arnsi du gène abcb6 humain et utilisation correspondante Ceased WO2019000148A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN2017/089928 WO2019000148A1 (fr) 2017-06-26 2017-06-26 Arnsi du gène abcb6 humain et utilisation correspondante

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2017/089928 WO2019000148A1 (fr) 2017-06-26 2017-06-26 Arnsi du gène abcb6 humain et utilisation correspondante

Publications (1)

Publication Number Publication Date
WO2019000148A1 true WO2019000148A1 (fr) 2019-01-03

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2017/089928 Ceased WO2019000148A1 (fr) 2017-06-26 2017-06-26 Arnsi du gène abcb6 humain et utilisation correspondante

Country Status (1)

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WO (1) WO2019000148A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116064527A (zh) * 2022-08-26 2023-05-05 广东省第二人民医院（广东省卫生应急医院） 抑制ABCB6基因表达的siRNA及其应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103849623A (zh) * 2012-11-28 2014-06-11 天津华大基因科技有限公司 Abcb6基因突变体及其应用

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103849623A (zh) * 2012-11-28 2014-06-11 天津华大基因科技有限公司 Abcb6基因突变体及其应用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE Nucleotide [O] 28 March 2017 (2017-03-28), ALLIKMETS, R. ET AL., XP055558442, retrieved from ncbi Database accession no. NM_005689 *
ZHAO, SHIGUANG ET AL.: "Increased Expression of ABCB6 Enhances Protoporphyrin IX Accumulation and Photodynamic Effect in Human Glioma", ANNALS OF SURGICAL ONCOLOGY, vol. 20, no. 13, 12 June 2012 (2012-06-12), pages 4379 - 4388, XP055558440, ISSN: 1068-9265 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116064527A (zh) * 2022-08-26 2023-05-05 广东省第二人民医院（广东省卫生应急医院） 抑制ABCB6基因表达的siRNA及其应用

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