Full metadata record
DC Field | Value | Language |
---|---|---|
dc.contributor.author | 고현철 | - |
dc.date.accessioned | 2019-12-09T05:54:24Z | - |
dc.date.available | 2019-12-09T05:54:24Z | - |
dc.date.issued | 2018-09 | - |
dc.identifier.citation | ARCHIVES OF PHARMACAL RESEARCH, v. 41, no. 9, page. 911-920 | en_US |
dc.identifier.issn | 0253-6269 | - |
dc.identifier.issn | 1976-3786 | - |
dc.identifier.uri | https://link.springer.com/article/10.1007%2Fs12272-018-1042-2 | - |
dc.identifier.uri | https://repository.hanyang.ac.kr/handle/20.500.11754/120096 | - |
dc.description.abstract | Targeted genome editing by clustered regularly interspaced short palindromic repeats (CRISPR-Cas9) raised concerns over off-target effects. The use of double-nicking strategy using paired Cas9 nickase has been developed to minimize off-target effects. However, it was reported that the efficiency of paired nickases were comparable or lower than that of either corresponding nuclease alone. Recently, we conducted a systematic comparison of the efficiencies of several paired Cas9 with their corresponding Cas9 nucleases and showed that paired D10A Cas9 nickases are sometimes more efficient than individual nucleases for gene disruption. However, sometimes the designed paired Cas9 nickases exhibited significantly lower mutation frequencies than nucleases, hampering the generation of cells containing paired Cas9 nickase-induced mutations. Here we implemented IRES peptide-conjugation of fluorescent protein to Cas9 nickase and subjected for fluorescence-activated cell sorting. The sorted cell populations are highly enriched with cells containing paired Cas9 nickase-induced mutations, by a factor of up to 40-fold as compared with the unsorted population. Furthermore, gene-disrupted single cell clones using paired nickases followed by FACS sorting strategy were generated highly efficiently, without compromising with its low off-target effects. We envision that our fluorescent protein coupled paired nickase-mediated gene disruption, facilitating efficient and highly specific genome editing in medical research. | en_US |
dc.description.sponsorship | This work was supported by a grant of Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health and Welfare, Republic of Korea (HI16C1009) and National Research Foundation of Korea (2017M3A9C6061361 and 2015R1C1A1A01054482). | en_US |
dc.language.iso | en_US | en_US |
dc.publisher | PHARMACEUTICAL SOC KOREA | en_US |
dc.subject | CRISPR-Cas9 | en_US |
dc.subject | IRES peptide-conjugation | en_US |
dc.subject | On-target efficiency | en_US |
dc.subject | Off-target efficiency | en_US |
dc.subject | Single cell clones | en_US |
dc.title | Efficient genome editing by FACS enrichment of paired D10A Cas9 nickases coupled with fluorescent proteins | en_US |
dc.type | Article | en_US |
dc.relation.no | 9 | - |
dc.relation.volume | 41 | - |
dc.identifier.doi | 10.1136/annrheumdis-2018-213328 | - |
dc.relation.page | 911-920 | - |
dc.relation.journal | ARCHIVES OF PHARMACAL RESEARCH | - |
dc.contributor.googleauthor | Gopalappa, Ramu | - |
dc.contributor.googleauthor | Song, Myungjae | - |
dc.contributor.googleauthor | Chandrasekaran, Arun Pandian | - |
dc.contributor.googleauthor | Das, Soumyadip | - |
dc.contributor.googleauthor | Haq, Saba | - |
dc.contributor.googleauthor | Koh, Hyun Chul | - |
dc.contributor.googleauthor | Ramakrishna, Suresh | - |
dc.relation.code | 2018007511 | - |
dc.sector.campus | S | - |
dc.sector.daehak | COLLEGE OF MEDICINE[S] | - |
dc.sector.department | DEPARTMENT OF MEDICINE | - |
dc.identifier.pid | hckoh | - |
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