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1. WO2022140232 - METHOD AND KIT FOR REGENERATING REUSABLE INITIATORS FOR NUCLEIC ACID SYNTHESIS

Publication Number WO/2022/140232
Publication Date 30.06.2022
International Application No. PCT/US2021/064298
International Filing Date 20.12.2021
IPC
C12P 19/34 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19Preparation of compounds containing saccharide radicals
26Preparation of nitrogen-containing carbohydrates
28N-glycosides
30Nucleotides
34Polynucleotides, e.g. nucleic acids, oligoribonucleotides
C12N 9/12 2006.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
10Transferases (2.)
12transferring phosphorus containing groups, e.g. kinases (2.7)
CPC
C12P 19/34
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19Preparation of compounds containing saccharide radicals
26Preparation of nitrogen-containing carbohydrates
28N-glycosides
30Nucleotides
34Polynucleotides, e.g. nucleic acids, oligoribonucleotides
Applicants
  • CHEN, Cheng-Yao
Inventors
  • CHEN, Cheng-Yao
  • CHENG, Yi-Wen
Agents
  • ROBERTS, Jonathan, A.
Priority Data
17/128,67721.12.2020US
Publication Language English (en)
Filing Language English (EN)
Designated States
Title
(EN) METHOD AND KIT FOR REGENERATING REUSABLE INITIATORS FOR NUCLEIC ACID SYNTHESIS
(FR) PROCÉDÉ ET KIT POUR RÉGÉNÉRER DES INITIATEURS RÉUTILISABLES POUR LA SYNTHÈSE D'ACIDE NUCLÉIQUE
Abstract
(EN) A method for nucleic acid synthesis and regeneration of a reusable synthesis initiator includes incorporating a linking nucleotide to an immobilized initiator using a polymerase, synthesizing a nucleic acid right after the linking nucleotide using the polymerase, subjecting a substrate base of the linking nucleotide in the nucleic acid to base-excision by a DNA glycosylase to generate an abasic site, subjecting the abasic site to cleavage by an endonuclease to release the nucleic acid from the initiator, and converting the 3' terminus of the initiator back to its original form by a 3' phosphatase activity-possessing enzyme. A kit based on the aforesaid method and a method tor regenerating a reusable initiator are also disclosed.
(FR) Procédé de synthèse d'acide nucléique et de régénération d'un initiateur de synthèse réutilisable comprenant les étapes suivantes : incorporation d'un nucléotide de liaison à un initiateur immobilisé en utilisant une polymérase; synthèse d'un acide nucléique juste après le nucléotide de liaison en utilisant la polymérase; soumission d'une base substrat du nucléotide de liaison dans l'acide nucléique à une excision de base par une ADN glycosylase pour générer un site abasique; soumission du site abasique à un clivage par une endonucléase pour libérer l'acide nucléique de l'initiateur; et conversion de l'extrémité 3' de l'initiateur en sa forme originale par une enzyme possédant une activité phosphatase 3'. L'invention concerne également un kit basé sur le procédé susmentionné et un procédé de régénération d'un initiateur réutilisable.
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