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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2020.6
- 수록면
- 114 - 119 (6page)
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초록· 키워드
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Of the various types of mice used for genome editing, conditional knock-out (cKO) mice serve as an important model for studying the function of genes. cKO mice can be produced using loxP knock-in (KI) mice in which loxP sequences (34 bp) are inserted on both sides of a specific region in the target gene. These mice can be used as KO mice that do not express a gene at a desired time or under a desired condition by cross-breeding with various Cre Tg mice. Genome editing has been recently made easy by the use of third-generation gene scissors, the CRISPR-Cas9 system. However, very few laboratories can produce mice for genome editing. Here we present a more efficient method for producing loxP KI mice. This method involves the use of an HDR vector as the target vector and ssODN as the donor DNA in order to induce homologous recombination for producing loxP KI mice. On injecting 20 ng/μL of ssODN, it was observed that the target exon was deleted or loxP was inserted on only one side. However, on injecting 10 ng/μL of the target HDR vector, the insertion of loxP was observed on both sides of the target region. In the first PCR, seven mice were identified to be loxP KI mice. The accuracy of their gene sequences was confirmed through Sanger sequencing. It is expected that the loxP KI mice produced in this study will serve as an important tool for identifying the function of the target gene.
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참고문헌 (22)
참고문헌 신청
Channabasavaiah B / 2019 / Creation of CRISPR-based germline-genome-engineered mice without ex vivo handling of zygotes by i-GONAD / Nat Prot 14:2452~2482
Cho SW / 2013 / Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease / Nat Biotechnol 31:230~232
Chu VT / 2015 / Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells / Nat Biotechnol 33:543~548
Chu VT / 2016 / Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6zygotes / BMC Biotechnol 16:86~97
Codner FG / 2018 / Application of long single-stranded DNA donors in genome editing : generation and validation of mouse mutants / BMC Biol 16:70
Cho SW / 2013 / Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease / Nat Biotechnol 31:230~232
Chu VT / 2015 / Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells / Nat Biotechnol 33:543~548
Chu VT / 2016 / Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6zygotes / BMC Biotechnol 16:86~97
Codner FG / 2018 / Application of long single-stranded DNA donors in genome editing : generation and validation of mouse mutants / BMC Biol 16:70
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UCI(KEPA) : I410-ECN-0101-2020-510-000849637