EP4720270A2 - Dnaps sujettes aux erreurs pour réplication d'adn orthogonale - Google Patents

Dnaps sujettes aux erreurs pour réplication d'adn orthogonale

Info

Publication number
EP4720270A2
EP4720270A2 EP24816418.8A EP24816418A EP4720270A2 EP 4720270 A2 EP4720270 A2 EP 4720270A2 EP 24816418 A EP24816418 A EP 24816418A EP 4720270 A2 EP4720270 A2 EP 4720270A2
Authority
EP
European Patent Office
Prior art keywords
mutation
sequences
trpb
selection
mutations
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.)
Pending
Application number
EP24816418.8A
Other languages
German (de)
English (en)
Inventor
Chang C. Liu
Gordon N. RIX
Rory L. WILLIAMS
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.)
University of California
University of California Berkeley
University of California San Diego UCSD
Original Assignee
University of California
University of California Berkeley
University of California San Diego UCSD
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 University of California, University of California Berkeley, University of California San Diego UCSD filed Critical University of California
Publication of EP4720270A2 publication Critical patent/EP4720270A2/fr
Pending legal-status Critical Current

Links

Classifications

    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
    • 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

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Medicinal Chemistry (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Une approche expérimentale accessible pour compresser une évolution génique à long terme à des échelles de temps de laboratoire permet l'évolution rapide de nouvelles fonctions biomoléculaires et pour l'observation directe d'une adaptation et d'une divergence étendues suivies par l'inférence de contraintes structurales, fonctionnelles et environnementales pour n'importe quel gène sélectionnable. L'invention concerne un système de réplication d'ADN orthogonale utilisant des ADN polymérases sujettes aux erreurs qui hypermute durablement des gènes choisis à une vitesse > 10-4 substitutions par base in vivo. Lorsqu'il est utilisé pour faire évoluer une enzyme maladaptée conditionnellement essentielle, une adaptation s'est produite rapidement et des milliers de séquences multi-mutations uniques ont été obtenues avec de nombreuses paires > 60 acides aminés à part (> 15 % de divergence), révélant des facteurs connus ainsi que des facteurs inattendus influençant l'adaptation enzymatique. L'aptitude des séquences évoluées n'a pas été prévisible par des modèles avancés d'apprentissage automatique entraînés sur une variation naturelle.
EP24816418.8A 2023-05-30 2024-05-30 Dnaps sujettes aux erreurs pour réplication d'adn orthogonale Pending EP4720270A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US202363504905P 2023-05-30 2023-05-30
PCT/US2024/031672 WO2024249634A2 (fr) 2023-05-30 2024-05-30 Dnaps sujettes aux erreurs pour réplication d'adn orthogonale

Publications (1)

Publication Number Publication Date
EP4720270A2 true EP4720270A2 (fr) 2026-04-08

Family

ID=93658783

Family Applications (1)

Application Number Title Priority Date Filing Date
EP24816418.8A Pending EP4720270A2 (fr) 2023-05-30 2024-05-30 Dnaps sujettes aux erreurs pour réplication d'adn orthogonale

Country Status (2)

Country Link
EP (1) EP4720270A2 (fr)
WO (1) WO2024249634A2 (fr)

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20230242902A1 (en) * 2017-10-20 2023-08-03 The Regents Of The University Of California A highly error-prone orthogonal dna replication system for targeted continuous evolution in vivo

Also Published As

Publication number Publication date
WO2024249634A3 (fr) 2025-04-17
WO2024249634A2 (fr) 2024-12-05

Similar Documents

Publication Publication Date Title
Paoli et al. Biosynthetic potential of the global ocean microbiome
Robinson et al. A roadmap for metagenomic enzyme discovery
Engqvist et al. Applications of protein engineering and directed evolution in plant research
Rix et al. Continuous evolution of user-defined genes at 1 million times the genomic mutation rate
Wanamaker et al. CrY2H-seq: a massively multiplexed assay for deep-coverage interactome mapping
US20210256394A1 (en) Methods and systems for the optimization of a biosynthetic pathway
Younger et al. Engineering modular biosensors to confer metabolite-responsive regulation of transcription
CA3029254A1 (fr) Procedes permettant de generer des bibliotheques combinatoires a code a barres
WO2012142591A2 (fr) Compositions, procédés et utilisations pour le mappage de la relation d'activité des séquences de protéines multiplexes
CN115349128A (zh) 宏基因组文库和天然产物发现平台
Belkhelfa et al. Continuous culture adaptation of Methylobacterium extorquens AM1 and TK 0001 to very high methanol concentrations
Mate et al. The pocket manual of directed evolution: tips and tricks
Qiao et al. Systematic characterization of hypothetical proteins in Synechocystis sp. PCC 6803 reveals proteins functionally relevant to stress responses
McClure et al. Species-specific transcriptomic network inference of interspecies interactions
de Pins et al. A systematic exploration of bacterial form I rubisco maximal carboxylation rates
Collins et al. Substrate-specific effects of natural genetic variation on proteasome activity
Pinto et al. Construction of a chassis for hydrogen production: physiological and molecular characterization of a Synechocystis sp. PCC 6803 mutant lacking a functional bidirectional hydrogenase
WO2024249634A2 (fr) Dnaps sujettes aux erreurs pour réplication d'adn orthogonale
US20190376067A1 (en) Compositions, methods and uses for multiplexed trackable genomically-engineered polypeptides
WO2024030344A1 (fr) Optimisation, basée sur un algorithme génétique et sur imodulon, d'une formulation de milieu pour des produits biologiques d'amélioration de qualité, de titre, de souche et de processus
Stikeleather et al. Translation Accuracy in E. coli
Jutur et al. Marine microalgae: exploring the systems through an omics approach for biofuel production
Rix Engineering in vivo hypermutation and selection systems for observing molecular evolution at scale
Hoffmann et al. The fitness landscape of a Form II rubisco in a photosynthetic bacterium guides engineering of oxygen tolerance
REPRODUCIBLE ESTABLISHING AFFINITY PURIFICATION-MASS SPECTROMETRY PROCEDURES PERMITTING REPRODUCIBLE AND HIGH-CONFIDENCE IDENTIFICATIONS OF IN VIVO PROTEIN INTERACTIONS 2

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20251215

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC ME MK MT NL NO PL PT RO RS SE SI SK SM TR