著作: [沢津橋 俊]/上甲 裕大/[福本 誠二]/[松本 俊夫]/Sugano S. Shigeo/Development of versatile non-homologous end joining-based knock-in module for genome editing./[Scientific Reports]
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種別 | 必須 | 学術論文(審査論文) | |||
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言語 | 必須 | 英語 | |||
招待 | 推奨 | ||||
審査 | 推奨 | ||||
カテゴリ | 推奨 | ||||
共著種別 | 推奨 | ||||
学究種別 | 推奨 | ||||
組織 | 推奨 | ||||
著者 | 必須 | ||||
題名 | 必須 |
(英) Development of versatile non-homologous end joining-based knock-in module for genome editing. |
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副題 | 任意 | ||||
要約 | 任意 |
(英) CRISPR/Cas9-based genome editing has dramatically accelerated genome engineering. An important aspect of genome engineering is efficient knock-in technology. For improved knock-in efficiency, the non-homologous end joining (NHEJ) repair pathway has been used over the homology-dependent repair pathway, but there remains a need to reduce the complexity of the preparation of donor vectors. We developed the versatile NHEJ-based knock-in module for genome editing (VIKING). Using the consensus sequence of the time-honored pUC vector to cut donor vectors, any vector with a pUC backbone could be used as the donor vector without customization. Conditions required to minimize random integration rates of the donor vector were also investigated. We attempted to isolate null lines of the VDR gene in human HaCaT keratinocytes using knock-in/knock-out with a selection marker cassette, and found 75% of clones isolated were successfully knocked-in. Although HaCaT cells have hypotetraploid genome composition, the results suggest multiple clones have VDR null phenotypes. VIKING modules enabled highly efficient knock-in of any vectors harboring pUC vectors. Users now can insert various existing vectors into an arbitrary locus in the genome. VIKING will contribute to low-cost genome engineering. |
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キーワード | 推奨 | ||||
発行所 | 推奨 | ||||
誌名 | 必須 |
Scientific Reports([Nature Publishing Group])
(eISSN: 2045-2322)
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巻 | 必須 | 8 | |||
号 | 必須 | 1 | |||
頁 | 必須 | 593 593 | |||
都市 | 任意 | ||||
年月日 | 必須 | 2018年 1月 12日 | |||
URL | 任意 | ||||
DOI | 任意 | 10.1038/s41598-017-18911-9 (→Scopusで検索) | |||
PMID | 任意 | 29330493 (→Scopusで検索) | |||
CRID | 任意 | ||||
WOS | 任意 | ||||
Scopus | 任意 | 2-s2.0-85043529637 | |||
機関リポジトリ | 112400 | ||||
評価値 | 任意 | ||||
被引用数 | 任意 | ||||
指導教員 | 推奨 | ||||
備考 | 任意 |
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