Title:
TECHNOLOGY FOR PREPARING NEW CLOSED LINEAR DOUBLE-STRANDED DNA IN VITRO
Document Type and Number:
WIPO Patent Application WO/2023/230991
Kind Code:
A1
Abstract:
A method for preparing a closed linear double-stranded DNA, comprising: (a) providing a double-stranded DNA comprising a target sequence and the cleavage sites of at least two restriction enzymes, wherein for each restriction enzyme, whose cleavage site is located between the recognition sequence thereof and the target sequence; (b) digesting the double-stranded DNA with restriction enzymes to generate a target DNA having non-palindromic sticky ends at both ends, wherein the two sticky ends cannot be complementarily paired with each other; (c) providing a stem-loop DNA having sticky ends complementarily paired with the sticky ends of the target DNA respectively, wherein the nucleotides in the loop of the stem-loop DNA are modified; and (d) linking the target DNA to the stem-loop DNA to form the closed linear double-stranded DNA.
Inventors:
ZHAO SHENG (CN)
TANG CHUANQING (CN)
TANG CHUANQING (CN)
Application Number:
PCT/CN2022/096863
Publication Date:
December 07, 2023
Filing Date:
June 02, 2022
Export Citation:
Assignee:
UBRIGENE SUZHOU BIOSCIENCES CO LTD (CN)
International Classes:
C12N15/09; C12N15/64; C12N15/10; C12Q1/02
Foreign References:
| US20040115616A1 | 2004-06-17 | |||
| CN101067133B | 2011-05-04 | |||
| CN107974448A | 2018-05-01 | |||
| CN108753807A | 2018-11-06 | |||
| US20110009281A1 | 2011-01-13 | |||
| US6451563B1 | 2002-09-17 |
Other References:
SCOTT VERONICA L, PATEL AMI, VILLARREAL DANIEL O, HENSLEY SCOTT E, RAGWAN EDWIN, YAN JIAN, SARDESAI NIRANJAN Y, ROTHWELL PAUL J, E: "Novel synthetic plasmid and Doggybone™ DNA vaccines induce neutralizing antibodies and provide protection from lethal influenza challenge in mice", HUMAN VACCINES & IMMUNOTHERAPEUTICS, TAYLOR & FRANCIS, US, vol. 11, no. 8, 3 August 2015 (2015-08-03), US , pages 1972 - 1982, XP093115901, ISSN: 2164-5515, DOI: 10.1080/21645515.2015.1022008
Attorney, Agent or Firm:
NTD UNIVATION INTELLECTUAL PROPERTY AGENCY LTD. (CN)
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