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CO2019000828A2 - Método para detectar un mutante dentro de una población de organismos aplicando el enfoque de agrupación y división - Google Patents

Método para detectar un mutante dentro de una población de organismos aplicando el enfoque de agrupación y división

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Publication number
CO2019000828A2
CO2019000828A2 CONC2019/0000828A CO2019000828A CO2019000828A2 CO 2019000828 A2 CO2019000828 A2 CO 2019000828A2 CO 2019000828 A CO2019000828 A CO 2019000828A CO 2019000828 A2 CO2019000828 A2 CO 2019000828A2
Authority
CO
Colombia
Prior art keywords
organisms
dpcr
finding
mutant
grouping
Prior art date
Application number
CONC2019/0000828A
Other languages
English (en)
Inventor
Toni Wendt
Ole Olsen
Søren Knudsen
Hanne Cecilie Thomsen
Alexander Striebeck
Birgitte Skadhauge
Magnus Wohlfahrt Rasmussen
Massimiliano Carciofi
Original Assignee
Carlsberg As
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 Carlsberg As filed Critical Carlsberg As
Publication of CO2019000828A2 publication Critical patent/CO2019000828A2/es

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    • 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
    • C12N15/01—Preparation of mutants without inserting foreign genetic material therein; Screening processes 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
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/46—Gramineae or Poaceae, e.g. ryegrass, rice, wheat or maize
    • A01H6/4624—Hordeum vulgarus [barley]
    • 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
    • C12N15/102—Mutagenizing nucleic acids
    • C12N15/1024—In vivo mutagenesis using high mutation rate "mutator" host strains by inserting genetic material, e.g. encoding an error prone polymerase, disrupting a gene for mismatch repair
    • 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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6858—Allele-specific amplification
    • 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
    • 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
    • C12Q2600/13—Plant traits
    • 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
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B30/00—Methods of screening libraries
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/02—Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biomedical Technology (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Plant Pathology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Developmental Biology & Embryology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Physiology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Cell Biology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

En los enfoques de mejoramiento de plantas tradicionales, puede utilizarse mutagénesis química para introducir sustituciones nucleotídicas aleatoriamente en el genoma de una planta, es decir sin posibilidades de controlar los sitios de los cambios nucleotídicos. A causa de las complejidades genómicas, la probabilidad estadística es extremadamente baja en cuanto a hallar una sustitución nucleotídica predeterminada. La presente invención, sin embargo, demuestra cómo se desarrolla un uso novedoso, alternativo, de la reacción en cadena de polimerasa digital (dPCR), preferiblemente dPCR en gotas (ddPCR), para explotar el hallazgo de sustituciones nucleotídicas específicas en genes mutados. La plataforma completa comprende un método de relevamiento con una biblioteca de organismos mutagenizados, sistemas basados en PCR digital y una configuración para propagar y analizar organismos mutados identificados.
CONC2019/0000828A 2016-07-01 2019-01-29 Método para detectar un mutante dentro de una población de organismos aplicando el enfoque de agrupación y división CO2019000828A2 (es)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DKPA201670485 2016-07-01
PCT/EP2017/065516 WO2018001884A1 (en) 2016-07-01 2017-06-23 Method to screen for a mutant within a population of organisms by applying a pooling and splitting approach

Publications (1)

Publication Number Publication Date
CO2019000828A2 true CO2019000828A2 (es) 2019-04-30

Family

ID=56681913

Family Applications (1)

Application Number Title Priority Date Filing Date
CONC2019/0000828A CO2019000828A2 (es) 2016-07-01 2019-01-29 Método para detectar un mutante dentro de una población de organismos aplicando el enfoque de agrupación y división

Country Status (25)

Country Link
US (2) US11884972B2 (es)
EP (3) EP3792357B1 (es)
JP (2) JP7208797B2 (es)
CN (2) CN114959098A (es)
AR (1) AR110103A1 (es)
AU (3) AU2017288960B2 (es)
BR (1) BR112018076429A2 (es)
CA (1) CA3026016A1 (es)
CL (1) CL2018003710A1 (es)
CO (1) CO2019000828A2 (es)
CR (1) CR20190040A (es)
CU (1) CU20180158A7 (es)
DK (1) DK3478832T3 (es)
EA (1) EA201892809A1 (es)
ES (1) ES2916221T3 (es)
IL (1) IL263451B (es)
MA (3) MA54568A (es)
MX (2) MX2018016110A (es)
PE (1) PE20190228A1 (es)
PL (2) PL3478832T3 (es)
PT (1) PT3478832T (es)
SG (1) SG11201811021YA (es)
UY (1) UY37312A (es)
WO (1) WO2018001884A1 (es)
ZA (3) ZA201808218B (es)

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MA54568A (fr) * 2016-07-01 2021-10-27 Carlsberg As Substitutions de nucléotides prédéterminées
US11690341B2 (en) 2017-12-28 2023-07-04 Carlsberg A/S Cereal plants with improved cell wall properties
CA3106307A1 (en) * 2018-07-13 2020-01-16 Hifibio Sas Use of droplet single cell epigenome profiling for patient stratification
US20220362771A1 (en) * 2018-07-13 2022-11-17 Hifibio Sas Use of droplet single cell epigenome profiling for patient stratification
CN108913698B (zh) * 2018-07-25 2021-10-26 安徽农业大学 一种与小麦穗发芽抗性/感性相关的caps标记及其应用
CN110724741B (zh) * 2019-07-24 2020-05-19 艾普拜生物科技(苏州)有限公司 一种检测微小残留白血病相关融合基因的引物、探针及其试剂盒
EP4013843A1 (en) * 2019-08-16 2022-06-22 Heineken Supply Chain B.V. Production of an alcohol-free beverage
EP3785529A1 (en) 2019-08-28 2021-03-03 Carlsberg A/S Barley plants with altered protein content in grains
CN114786473B (zh) * 2019-10-10 2023-12-19 嘉士伯有限公司 制备突变植物的方法
WO2021175786A1 (en) 2020-03-02 2021-09-10 Carlsberg A/S Barley plants with high limit dextrinase activity
EP4029935A1 (en) 2021-01-14 2022-07-20 Carlsberg A/S S. eubayanus strain and hybrids thereof
JP2024538973A (ja) * 2021-10-25 2024-10-28 ジーティー モレキュラー インコーポレイテッド ポリメラーゼ連鎖反応によるターゲット識別を向上させるオフターゲットブロック配列
EP4451853A1 (en) 2021-12-22 2024-10-30 Basf Agricultural Solutions Seed Us Llc Improved screening method for genome edited events
EP4490736A1 (en) 2022-03-11 2025-01-15 Carlsberg A/S Modulation of protein levels
WO2024105148A1 (en) 2022-11-16 2024-05-23 University Of Copenhagen Plants with low seed glucosinolate content
WO2024133900A1 (en) 2022-12-22 2024-06-27 University Of Copenhagen Improved (-)-sparteine production
WO2024227748A1 (en) 2023-05-01 2024-11-07 Carlsberg A/S Barley with improved properties
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CN117965576B (zh) * 2024-02-01 2024-08-27 北京林业大学 一种王百合苷生物合成基因LlHCT1及其应用

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CA3026016A1 (en) 2018-01-04
MX2018016110A (es) 2019-05-30
PL3792357T3 (pl) 2025-07-07
EP3478832B1 (en) 2022-02-23
CN109477087B (zh) 2022-06-24
AR110103A1 (es) 2019-02-27
MX2022014965A (es) 2023-01-11
CN109477087A (zh) 2019-03-15
MA52594A (fr) 2021-03-17
DK3478832T3 (da) 2022-05-30
IL263451B (en) 2022-08-01
JP7539428B2 (ja) 2024-08-23
EP3693458A1 (en) 2020-08-12
AU2017288960A1 (en) 2019-01-17
EP3478832A1 (en) 2019-05-08
MA54568A (fr) 2021-10-27
US20240182961A1 (en) 2024-06-06
AU2019268102B2 (en) 2020-03-05
BR112018076429A2 (pt) 2019-10-01
AU2019268103A1 (en) 2019-12-12
UY37312A (es) 2018-02-28
ZA201907775B (en) 2022-09-28
CR20190040A (es) 2019-05-09
PE20190228A1 (es) 2019-02-13
EA201892809A1 (ru) 2019-06-28
IL263451A (en) 2019-01-31
AU2019268103B2 (en) 2020-03-12
PT3478832T (pt) 2022-05-19
JP2019524083A (ja) 2019-09-05
SG11201811021YA (en) 2019-01-30
EP3792357C0 (en) 2025-05-07
CN114959098A (zh) 2022-08-30
WO2018001884A1 (en) 2018-01-04
AU2017288960B2 (en) 2019-12-12
CL2018003710A1 (es) 2019-03-22
AU2019268102A1 (en) 2019-12-12
CU20180158A7 (es) 2019-08-06
US20190194723A1 (en) 2019-06-27
JP2022101703A (ja) 2022-07-06
EP3792357A1 (en) 2021-03-17
EP3792357B1 (en) 2025-05-07
US11884972B2 (en) 2024-01-30
ES2916221T3 (es) 2022-06-29
ZA201907774B (en) 2022-07-27
ZA201808218B (en) 2020-02-26
MA45531A (fr) 2019-05-08
JP7208797B2 (ja) 2023-01-19
PL3478832T3 (pl) 2022-06-06

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