Cited 9 times in
High Homology-Directed Repair Using Mitosis Phase and Nucleus Localizing Signal
DC Field | Value | Language |
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dc.date.accessioned | 2022-09-02T01:16:16Z | - |
dc.date.available | 2022-09-02T01:16:16Z | - |
dc.date.issued | 2020-05 | - |
dc.identifier.issn | 1661-6596 | - |
dc.identifier.uri | https://ir.ymlib.yonsei.ac.kr/handle/22282913/190107 | - |
dc.description.abstract | In homology-directed repair, mediated knock-in single-stranded oligodeoxynucleotides (ssODNs) can be used as a homologous template and present high efficiency, but there is still a need to improve efficiency. Previous studies have mainly focused on controlling double-stranded break size, ssODN stability, and the DNA repair cycle. Nevertheless, there is a lack of research on the correlation between the cell cycle and single-strand template repair (SSTR) efficiency. Here, we investigated the relationship between cell cycle and SSTR efficiency. We found higher SSTR efficiency during mitosis, especially in the metaphase and anaphase. A Cas9 protein with a nuclear localization signal (NLS) readily migrated to the nucleus; however, the nuclear envelope inhibited the nuclear import of many nucleotide templates. This seemed to result in non-homologous end joining (NHEJ) before the arrival of the homologous template. Thus, we assessed whether NLS-tagged ssODNs and free NLS peptides could circumvent problems posed by the nuclear envelope. NLS-tagging ssODNs enhanced SSTR and indel efficiency by 4-fold compared to the control. Our results suggest the following: (1) mitosis is the optimal phase for SSTR, (2) the donor template needs to be delivered to the nucleus before nuclease delivery, and (3) NLS-tagging ssODNs improve SSTR efficiency, especially high in mitosis. | - |
dc.description.statementOfResponsibility | open | - |
dc.language | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | - |
dc.publisher | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | - |
dc.relation.isPartOf | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | - |
dc.rights | CC BY-NC-ND 2.0 KR | - |
dc.subject.MESH | Animals | - |
dc.subject.MESH | CRISPR-Associated Protein 9 / genetics* | - |
dc.subject.MESH | CRISPR-Associated Protein 9 / metabolism | - |
dc.subject.MESH | CRISPR-Cas Systems | - |
dc.subject.MESH | Gene Editing / methods* | - |
dc.subject.MESH | Gene Knock-In Techniques / methods* | - |
dc.subject.MESH | Mice | - |
dc.subject.MESH | Mice, Inbred C57BL | - |
dc.subject.MESH | Mitosis* | - |
dc.subject.MESH | NIH 3T3 Cells | - |
dc.subject.MESH | Nuclear Localization Signals* | - |
dc.subject.MESH | Recombinant Proteins / genetics | - |
dc.subject.MESH | Recombinant Proteins / metabolism | - |
dc.subject.MESH | Recombinational DNA Repair | - |
dc.title | High Homology-Directed Repair Using Mitosis Phase and Nucleus Localizing Signal | - |
dc.type | Article | - |
dc.contributor.college | College of Medicine (의과대학) | - |
dc.contributor.department | Dept. of Pharmacology (약리학교실) | - |
dc.contributor.googleauthor | Jeong Pil Han | - |
dc.contributor.googleauthor | Yoo Jin Chang | - |
dc.contributor.googleauthor | Dong Woo Song | - |
dc.contributor.googleauthor | Beom Seok Choi | - |
dc.contributor.googleauthor | Ok Jae Koo | - |
dc.contributor.googleauthor | Seung Youn Yi | - |
dc.contributor.googleauthor | Tae Sub Park | - |
dc.contributor.googleauthor | Su Cheong Yeom | - |
dc.identifier.doi | 10.3390/ijms21113747 | - |
dc.relation.journalcode | J01133 | - |
dc.identifier.eissn | 1422-0067 | - |
dc.identifier.pmid | 32466470 | - |
dc.subject.keyword | CRISPR | - |
dc.subject.keyword | embryo | - |
dc.subject.keyword | homology-directed repair | - |
dc.subject.keyword | mitosis | - |
dc.subject.keyword | NLS | - |
dc.subject.keyword | ssODN | - |
dc.citation.volume | 21 | - |
dc.citation.number | 11 | - |
dc.citation.startPage | 3747 | - |
dc.identifier.bibliographicCitation | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, Vol.21(11) : 3747, 2020-05 | - |
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