Title:
METHOD FOR PRODUCING TEMPLATE DNA
Document Type and Number:
WIPO Patent Application WO/2023/085117
Kind Code:
A1
Abstract:
This method for producing a template DNA comprises: step (a) for preparing a cassette double-stranded DNA; step (b) for preparing an oligonucleotide containing a target sequence; step (c) for ligating the same by a DNA ligase to thereby obtain a cyclic nucleotide containing the target sequence; step (d) for, using the nucleotide as a template, amplifying by a strand displacement DNA polymerase to thereby obtain a double-stranded DNA containing the target sequence that is a concatemer having the same sequences consisting of a support sequence, an RNA polymerase promoter sequence, the target sequence and an sgRNA scaffold sequence; and step (e) for cleaving the double-stranded DNA between the same sequences to thereby obtain a template DNA having sequences consisting of the support sequence, the RNA polymerase promoter sequence, the target sequence and the sgRNA scaffold sequence.
Inventors:
OKAMURA YOSHIKO (JP)
TAKAHASHI HIROKAZU (JP)
TAKAHASHI HIROKAZU (JP)
Application Number:
PCT/JP2022/040284
Publication Date:
May 19, 2023
Filing Date:
October 28, 2022
Export Citation:
Assignee:
UNIV HIROSHIMA (JP)
International Classes:
C12N15/09; C12Q1/6806
Foreign References:
| JP2010051182A | 2010-03-11 |
Other References:
KELLEY MELISSA L.; STREZOSKA ŽAKLINA; HE KAIZHANG; VERMEULEN ANNALEEN; SMITH ANJA VAN BRABANT: "Versatility of chemically synthesized guide RNAs for CRISPR-Cas9 genome editing", JOURNAL OF BIOTECHNOLOGY, ELSEVIER, AMSTERDAM NL, vol. 233, 30 June 2016 (2016-06-30), Amsterdam NL , pages 74 - 83, XP029674969, ISSN: 0168-1656, DOI: 10.1016/j.jbiotec.2016.06.011
Attorney, Agent or Firm:
ISEKI Katsumori et al. (JP)
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