GB2631830B - Linear DNA with enhanced resistance against exonucleases and methods for the production thereof - Google Patents

Linear DNA with enhanced resistance against exonucleases and methods for the production thereof

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Publication number
GB2631830B
GB2631830B GB2407102.9A GB202407102A GB2631830B GB 2631830 B GB2631830 B GB 2631830B GB 202407102 A GB202407102 A GB 202407102A GB 2631830 B GB2631830 B GB 2631830B
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GB
United Kingdom
Prior art keywords
production
methods
resistance against
linear dna
enhanced resistance
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.)
Active
Application number
GB2407102.9A
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English (en)
Other versions
GB2631830A (en
GB202407102D0 (en
Inventor
Picher Ángel
Lanckriet Heikki
Walker Amy
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.)
4Basebio SL
4Basebio UK Ltd
Original Assignee
4Basebio SL
4Basebio UK Ltd
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Priority to GB2508033.4A priority Critical patent/GB2700069A/en
Publication of GB202407102D0 publication Critical patent/GB202407102D0/en
Publication of GB2631830A publication Critical patent/GB2631830A/en
Application granted granted Critical
Publication of GB2631830B publication Critical patent/GB2631830B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

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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
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • 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
    • C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
    • C12Q1/6855—Ligating adaptors
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • 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
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/12—Type of nucleic acid catalytic nucleic acids, e.g. ribozymes
    • C12N2310/122—Hairpin
    • C—CHEMISTRY; METALLURGY
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    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/30—Chemical structure
    • C12N2310/31—Chemical structure of the backbone
    • C12N2310/315—Phosphorothioates
    • C—CHEMISTRY; METALLURGY
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    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • 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
    • C12Q2521/00—Reaction characterised by the enzymatic activity
    • C12Q2521/50—Other enzymatic activities
    • C12Q2521/501—Ligase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/10—Modifications characterised by
    • C12Q2525/125—Modifications characterised by incorporating agents resulting in resistance to degradation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/10—Modifications characterised by
    • C12Q2525/191—Modifications characterised by incorporating an adaptor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/30—Oligonucleotides characterised by their secondary structure
    • C12Q2525/301—Hairpin oligonucleotides
    • 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
    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/30—Oligonucleotides characterised by their secondary structure
    • C12Q2525/307—Circular oligonucleotides
    • 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
    • C12Q2531/00—Reactions of nucleic acids characterised by
    • C12Q2531/10—Reactions of nucleic acids characterised by the purpose being amplify/increase the copy number of target nucleic acid
    • C12Q2531/125—Rolling circle
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2537/00—Reactions characterised by the reaction format or use of a specific feature
    • C12Q2537/10—Reactions characterised by the reaction format or use of a specific feature the purpose or use of
    • C12Q2537/101—Homogeneous assay format, e.g. one pot reaction

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Plant Pathology (AREA)
  • Immunology (AREA)
  • Cell Biology (AREA)
  • Virology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
GB2407102.9A 2021-07-30 2022-07-29 Linear DNA with enhanced resistance against exonucleases and methods for the production thereof Active GB2631830B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
GB2508033.4A GB2700069A (en) 2021-07-30 2022-07-29 Linear dna with enhanced resistance against exonucleases and methods for the production thereof

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP21382725.6A EP4124660A1 (en) 2021-07-30 2021-07-30 Linear dna with enhanced resistance against exonucleases and methods for the production thereof
GB2317071.5A GB2621500B (en) 2021-07-30 2022-07-29 Methods for the production of linear DNA with enhanced resistance against exonucleases

Publications (3)

Publication Number Publication Date
GB202407102D0 GB202407102D0 (en) 2024-07-03
GB2631830A GB2631830A (en) 2025-01-15
GB2631830B true GB2631830B (en) 2025-10-08

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ID=77168191

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GB2407102.9A Active GB2631830B (en) 2021-07-30 2022-07-29 Linear DNA with enhanced resistance against exonucleases and methods for the production thereof
GB2508033.4A Pending GB2700069A (en) 2021-07-30 2022-07-29 Linear dna with enhanced resistance against exonucleases and methods for the production thereof
GB2317071.5A Active GB2621500B (en) 2021-07-30 2022-07-29 Methods for the production of linear DNA with enhanced resistance against exonucleases

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GB2508033.4A Pending GB2700069A (en) 2021-07-30 2022-07-29 Linear dna with enhanced resistance against exonucleases and methods for the production thereof
GB2317071.5A Active GB2621500B (en) 2021-07-30 2022-07-29 Methods for the production of linear DNA with enhanced resistance against exonucleases

Country Status (11)

Country Link
US (1) US20240076659A1 (cg-RX-API-DMAC7.html)
EP (1) EP4124660A1 (cg-RX-API-DMAC7.html)
JP (1) JP2024528060A (cg-RX-API-DMAC7.html)
KR (1) KR20240038989A (cg-RX-API-DMAC7.html)
CN (1) CN117980501A (cg-RX-API-DMAC7.html)
AU (1) AU2022320939A1 (cg-RX-API-DMAC7.html)
CA (1) CA3224561A1 (cg-RX-API-DMAC7.html)
ES (3) ES2947031T6 (cg-RX-API-DMAC7.html)
GB (3) GB2631830B (cg-RX-API-DMAC7.html)
MX (1) MX2024001110A (cg-RX-API-DMAC7.html)
WO (1) WO2023006978A1 (cg-RX-API-DMAC7.html)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
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EP4553156A1 (en) 2023-11-08 2025-05-14 4basebio UK Ltd Synthetic dna production using cas nuclease
WO2025120370A1 (en) * 2023-12-08 2025-06-12 Camena Bioscience Limited Compositions and methods for solution-phase, phosphoramidite-free synthesis of nucleic acids
EP4596713A1 (en) 2024-02-05 2025-08-06 4basebio UK Ltd Dna with enhanced resistance against exonucleases and methods for the production thereof
EP4628594A1 (en) 2024-04-05 2025-10-08 4basebio UK Ltd Methods of amplifying dna and dna products with inhibitory regions
WO2025235462A1 (en) * 2024-05-06 2025-11-13 Gt Molecular, Inc. Selective blocking to detect and amplify low abundant template
WO2025238067A1 (en) 2024-05-15 2025-11-20 Ascend Advanced Therapies Gmbh Novel vector system for aav production

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WO2022058755A1 (en) * 2020-09-18 2022-03-24 Lightbio Limited Self-targeting expression vector
WO2023006985A1 (en) * 2021-07-30 2023-02-02 4Basebio Uk Ltd Nanoparticles and peptides for the delivery of cargos to muscle cells

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KR101414713B1 (ko) * 2007-10-11 2014-07-03 삼성전자주식회사 리가제 및 엔도뉴클레아제의 존재하에서 롤링서클 증폭에의하여 표적 핵산을 증폭하는 방법
GB0901593D0 (en) 2009-01-30 2009-03-11 Touchlight Genetics Ltd Production of closed linear DNA
US20120238738A1 (en) * 2010-07-19 2012-09-20 New England Biolabs, Inc. Oligonucleotide Adapters: Compositions and Methods of Use
WO2018031588A1 (en) * 2016-08-09 2018-02-15 Takara Bio Usa, Inc. Nucleic acid adaptors with molecular identification sequences and use thereof
US10350307B2 (en) * 2017-09-18 2019-07-16 General Electric Company In vivo RNA or protein expression using double-stranded concatemeric DNA including phosphorothioated nucleotides
CN114616338A (zh) * 2019-08-22 2022-06-10 新加坡国立大学 生成哑铃形dna载体的方法
TWI771847B (zh) * 2019-12-30 2022-07-21 財團法人工業技術研究院 擴增和確定目標核苷酸序列的方法
AU2021213927A1 (en) * 2020-01-31 2022-07-28 Tyris Therapeutics, S.L. Closed linear DNA with modified nucleotides
AU2021212319A1 (en) * 2020-01-31 2022-07-28 Tyris Therapeutics, S.L. Process for the production of closed linear DNA
WO2023230991A1 (zh) * 2022-06-02 2023-12-07 宜明(苏州)细胞生物科技有限公司 新型闭合线性双链dna的体外制备技术

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WO2022058755A1 (en) * 2020-09-18 2022-03-24 Lightbio Limited Self-targeting expression vector
WO2023006985A1 (en) * 2021-07-30 2023-02-02 4Basebio Uk Ltd Nanoparticles and peptides for the delivery of cargos to muscle cells

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Also Published As

Publication number Publication date
ES2956243A2 (es) 2023-12-15
KR20240038989A (ko) 2024-03-26
GB2631830A (en) 2025-01-15
US20240076659A1 (en) 2024-03-07
ES2996704A2 (es) 2025-02-13
ES2947031T6 (cg-RX-API-DMAC7.html) 2023-08-02
GB2700069A (en) 2025-09-10
WO2023006978A1 (en) 2023-02-02
GB2621500B (en) 2024-07-03
GB202317071D0 (en) 2023-12-20
CN117980501A (zh) 2024-05-03
GB202508033D0 (en) 2025-07-09
MX2024001110A (es) 2024-05-09
CA3224561A1 (en) 2023-02-02
GB202407102D0 (en) 2024-07-03
ES2956243B2 (es) 2024-11-26
JP2024528060A (ja) 2024-07-26
EP4124660A1 (en) 2023-02-01
AU2022320939A1 (en) 2024-02-29
ES2996704R1 (es) 2025-07-16
GB2621500A (en) 2024-02-14
ES2956243R1 (es) 2024-08-07

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