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CN111278983A - 基因敲除方法 - Google Patents

基因敲除方法 Download PDF

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Publication number
CN111278983A
CN111278983A CN201780093414.5A CN201780093414A CN111278983A CN 111278983 A CN111278983 A CN 111278983A CN 201780093414 A CN201780093414 A CN 201780093414A CN 111278983 A CN111278983 A CN 111278983A
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CN
China
Prior art keywords
target sequence
universal
donor dna
cell
linear donor
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Pending
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CN201780093414.5A
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English (en)
Inventor
魏文胜
陈一欧
周悦欣
张鸿敏
袁鹏飞
刘源
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Peking University
Edigene Biotechnology Inc
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Peking University
Edigene Biotechnology Inc
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Publication of CN111278983A publication Critical patent/CN111278983A/zh
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    • C12N15/09—Recombinant DNA-technology
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
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    • 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
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
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    • 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
    • C12N15/65—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression using markers
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
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    • G01—MEASURING; TESTING
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    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPR]
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/35—Nature of the modification
    • C12N2310/351—Conjugate
    • C12N2310/3517—Marker; Tag

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  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

提供一种供体构建体，以及基因敲除方法及其系统和试剂盒。所述体构建体是线性供体DNA或可在细胞中被切割产生线性供体DNA。所述基因敲除方法利用供体构建体中包含的标记物基因富集被基因敲除的细胞，提高通过序列特异性核酸酶产生基因敲除的效率。

Description

PCT国内申请，说明书已公开。

Claims (10)

  1. PCT国内申请，权利要求书已公开。
CN201780093414.5A 2017-08-08 2017-08-08 基因敲除方法 Pending CN111278983A (zh)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN2017/096510 WO2019028686A1 (zh) 2017-08-08 2017-08-08 基因敲除方法

Publications (1)

Publication Number Publication Date
CN111278983A true CN111278983A (zh) 2020-06-12

Family

ID=65273132

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201780093414.5A Pending CN111278983A (zh) 2017-08-08 2017-08-08 基因敲除方法

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Country Link
US (1) US11624077B2 (zh)
EP (1) EP3666898A4 (zh)
JP (1) JP7109009B2 (zh)
CN (1) CN111278983A (zh)
WO (1) WO2019028686A1 (zh)

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WO2019024081A1 (zh) 2017-08-04 2019-02-07 北京大学 特异性识别甲基化修饰dna碱基的tale rvd及其应用
CN113980999A (zh) * 2021-01-21 2022-01-28 扬州大学 基于多位点同时敲除的CRISPR/Cas9载体及其构建方法和应用

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Also Published As

Publication number Publication date
US20200255866A1 (en) 2020-08-13
US11624077B2 (en) 2023-04-11
EP3666898A1 (en) 2020-06-17
EP3666898A4 (en) 2021-03-24
JP2020533017A (ja) 2020-11-19
JP7109009B2 (ja) 2022-07-29
WO2019028686A1 (zh) 2019-02-14

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Application publication date: 20200612