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1. WO2022126304 - MODIFIED HELICASE AND APPLICATION THEREOF

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Publication Number WO/2022/126304
Publication Date 23.06.2022
International Application No. PCT/CN2020/136031
International Filing Date 14.12.2020
IPC
C12N 9/14 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
14Hydrolases (3.)
C12N 15/55 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
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
52Genes encoding for enzymes or proenzymes
55Hydrolases (3)
C12Q 1/6869 2018.1
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
6869Methods for sequencing
G01N 33/48 2006.1
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
Applicants
  • 北京齐碳科技有限公司 QITAN TECHNOLOGY LTD., BEIJING [CN]/[CN]
Inventors
  • 王慕旸 WANG, MuYang
  • 张周刚 ZHANG, Zhougang
  • 吕伟丽 LV, Weili
  • 王艳双 WANG, Yanshuang
  • 陈呈尧 CHEN, Chengyao
Agents
  • 北京领科知识产权代理事务所（特殊普通合伙） BEIJING LINKTEC IP LAW FIRM (SPECIAL GENERAL PARTNERSHIP)
Priority Data
Publication Language Chinese (zh)
Filing Language Chinese (ZH)
Designated States
Title
(EN) MODIFIED HELICASE AND APPLICATION THEREOF
(FR) HÉLICASE MODIFIÉE ET SON APPLICATION
(ZH) 一种经修饰的解旋酶及其应用
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Abstract
(EN) Provided are a modified F8813 helicase, a construct comprising the modified F8813 helicase, and an application in characterizing a target polynucleotide or controlling the movement of a target polynucleotide passing through pores. Also provided are a method for controlling the movement of a polynucleotide or a method for characterizing the target polynucleotide. The modified F8813 helicase can be used to control the translocation of a DNA chain through transmembrane pores in DNA chain sequencing, and also can realize direct RNA sequencing. Moreover, the modified helicase can remain bound to the polynucleotide for a longer time, thereby reducing the phenomenon of being released from a polynucleotide of a sequenced polynucleotide.
(FR) La présente invention concerne une hélicase F8813 modifiée, une construction comprenant l'hélicase F8813 modifiée, et une application dans la caractérisation d'un polynucléotide cible ou la régulation du mouvement d'un polynucléotide cible traversant des pores. L'invention concerne également un procédé de régulation du mouvement d'un polynucléotide ou un procédé de caractérisation du polynucléotide cible. L'hélicase F8813 modifiée peut être utilisée pour réguler la translocation d'une chaîne d'ADN à travers les pores transmembranaires dans le séquençage de la chaîne d'ADN, et peut également réaliser le séquençage direct de l'ARN. De plus, l'hélicase modifiée peut rester liée au polynucléotide pendant un temps plus long, réduisant ainsi le phénomène de libération d'un polynucléotide d'un polynucléotide séquencé.
(ZH) 提供了一种经修饰的F8813解旋酶，包含经修饰的F8813解旋酶的构建体，以及在表征目标多核苷酸或控制目标多核苷酸穿过孔的移动中的应用。还提供了一种控制多核苷酸移动的方法或一种表征目标多核苷酸的方法。采用经修饰的F8813解旋酶可以实现在DNA链测序中控制其穿过跨膜孔的移位，也可以实现对RNA的直接测序，而且，该经修饰的解旋酶可以更长时间保持与多核苷酸的结合，降低从被测序的多核苷酸上解脱的现象。
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