WEKO3
アイテム
{"_buckets": {"deposit": "98c7fd8c-4623-433f-8e9e-7c95f6216564"}, "_deposit": {"id": "41671", "owners": [], "pid": {"revision_id": 0, "type": "depid", "value": "41671"}, "status": "published"}, "_oai": {"id": "oai:tsukuba.repo.nii.ac.jp:00041671", "sets": ["918", "1782", "5975"]}, "item_5_biblio_info_6": {"attribute_name": "書誌情報", "attribute_value_mlt": [{"bibliographicIssueDates": {"bibliographicIssueDate": "2017-05", "bibliographicIssueDateType": "Issued"}, "bibliographicIssueNumber": "1", "bibliographicPageEnd": "18", "bibliographicPageStart": "14", "bibliographicVolumeNumber": "26", "bibliographic_titles": [{"bibliographic_title": "Medical journal of Indonesia"}]}]}, "item_5_creator_3": {"attribute_name": "著者別名", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "渡邊, 幸秀"}], "nameIdentifiers": [{"nameIdentifier": "447", "nameIdentifierScheme": "WEKO"}, {"nameIdentifier": "40618534", "nameIdentifierScheme": "e-Rad", "nameIdentifierURI": "https://nrid.nii.ac.jp/ja/nrid/1000040618534"}, {"nameIdentifier": "0000003055", "nameIdentifierScheme": "筑波大学研究者総覧", "nameIdentifierURI": "http://trios.tsukuba.ac.jp/researcher/0000003055"}]}, {"creatorNames": [{"creatorName": "加藤, 光保"}], "nameIdentifiers": [{"nameIdentifier": "611", "nameIdentifierScheme": "WEKO"}, {"nameIdentifier": "20194855", "nameIdentifierScheme": "e-Rad", "nameIdentifierURI": "https://nrid.nii.ac.jp/ja/nrid/1000020194855"}, {"nameIdentifier": "0000001657", "nameIdentifierScheme": "筑波大学研究者総覧", "nameIdentifierURI": "http://trios.tsukuba.ac.jp/researcher/0000001657"}]}]}, "item_5_description_4": {"attribute_name": "抄録", "attribute_value_mlt": [{"subitem_description": "Background:\n Clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9) is a powerful genome editing technique. It consists of RNA-guided DNA endonuclease Cas9 and single guide RNA (gRNA). By combining their expressions, high efficiency cleavage of the target gene can be achieved, leading to the formation of DNA double-strand break (DSB) at the genomic locus of interest which will be repaired via NHEJ (non-homologous end joining) or HDR (homology-directed repair) and mediate DNA alteration. We aimed to apply the CRISPR/Cas9 technique to knock-out the transmembrane prostate androgen-induced protein (TMEPAI) gene in the triple negative breast cancer cell line.\nMethods:\n Designed gRNA which targets the TMEPAI gene was synthesized, annealed, and cloned into gRNA expression vector. It was co-transfected into the TNBC cell line using polyethylenimine (PEI) together with Cas9-GFP and puromycin resistant gene vector. At 24-hours post-transfection, cells were selected by puromycin for 3 days before they were cloned. Selected knock-out clones were subsequently checked on their protein levels by western blotting.\nResults:\nCRISPR/Cas9, a genome engineering technique successfully knocked-out TMEPAI in the Hs578T TNBC cell line. Sequencing shows a frameshift mutation in TMEPAI. Western blot shows the absence of TMEPAI band on Hs578T KO cells.\nConclusion: \nTMEPAI gene was deleted in the TNBC cell line using the genomic editing technique CRISPR/Cas9. The deletion was confirmed by genome and protein analysis.", "subitem_description_type": "Abstract"}]}, "item_5_publisher_27": {"attribute_name": "出版者", "attribute_value_mlt": [{"subitem_publisher": "University of Indonesia, Faculty of Medicine"}]}, "item_5_relation_11": {"attribute_name": "DOI", "attribute_value_mlt": [{"subitem_relation_type_id": {"subitem_relation_type_id_text": "10.13181/mji.v26i1.1871", "subitem_relation_type_select": "DOI"}}]}, "item_5_rights_12": {"attribute_name": "権利", "attribute_value_mlt": [{"subitem_rights": "Copyright @ 2017 Authors."}, {"subitem_rights": "This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original author and source are properly cited."}]}, "item_5_select_15": {"attribute_name": "著者版フラグ", "attribute_value_mlt": [{"subitem_select_item": "publisher"}]}, "item_5_source_id_7": {"attribute_name": "ISSN", "attribute_value_mlt": [{"subitem_source_identifier": "2252-8083", "subitem_source_identifier_type": "ISSN"}]}, "item_5_source_id_9": {"attribute_name": "書誌レコードID", "attribute_value_mlt": [{"subitem_source_identifier": "AA1113872X", "subitem_source_identifier_type": "NCID"}]}, "item_creator": {"attribute_name": "著者", "attribute_type": "creator", "attribute_value_mlt": [{"creatorNames": [{"creatorName": "Wardhani, Bantari W.K."}], "nameIdentifiers": [{"nameIdentifier": "164768", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Puteri, Meidi U."}], "nameIdentifiers": [{"nameIdentifier": "164769", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Watanabe, Yukihide"}], "nameIdentifiers": [{"nameIdentifier": "164770", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Louisa, Melva"}], "nameIdentifiers": [{"nameIdentifier": "164771", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Setiabudy, Rianto"}], "nameIdentifiers": [{"nameIdentifier": "164772", "nameIdentifierScheme": "WEKO"}]}, {"creatorNames": [{"creatorName": "Kato, Mitsuyasu"}], "nameIdentifiers": [{"nameIdentifier": "164773", "nameIdentifierScheme": "WEKO"}]}]}, "item_files": {"attribute_name": "ファイル情報", "attribute_type": "file", "attribute_value_mlt": [{"accessrole": "open_date", "date": [{"dateType": "Available", "dateValue": "2017-06-23"}], "displaytype": "detail", "download_preview_message": "", "file_order": 0, "filename": "MJI_26-1.pdf", "filesize": [{"value": "566.3 kB"}], "format": "application/pdf", "future_date_message": "", "is_thumbnail": false, "licensetype": "license_9", "mimetype": "application/pdf", "size": 566300.0, "url": {"label": "MJI_26-1", "url": "https://tsukuba.repo.nii.ac.jp/record/41671/files/MJI_26-1.pdf"}, "version_id": "faf3e71f-b592-4d06-831c-cd7dd088011d"}]}, "item_language": {"attribute_name": "言語", "attribute_value_mlt": [{"subitem_language": "eng"}]}, "item_resource_type": {"attribute_name": "資源タイプ", "attribute_value_mlt": [{"resourcetype": "journal article", "resourceuri": "http://purl.org/coar/resource_type/c_6501"}]}, "item_title": "TMEPAI genome editing in triple negative breast cancer cells", "item_titles": {"attribute_name": "タイトル", "attribute_value_mlt": [{"subitem_title": "TMEPAI genome editing in triple negative breast cancer cells"}]}, "item_type_id": "5", "owner": "1", "path": ["918", "1782", "5975"], "permalink_uri": "http://hdl.handle.net/2241/00146612", "pubdate": {"attribute_name": "公開日", "attribute_value": "2017-06-23"}, "publish_date": "2017-06-23", "publish_status": "0", "recid": "41671", "relation": {}, "relation_version_is_last": true, "title": ["TMEPAI genome editing in triple negative breast cancer cells"], "weko_shared_id": 5}
TMEPAI genome editing in triple negative breast cancer cells
http://hdl.handle.net/2241/00146612
http://hdl.handle.net/2241/001466122a0ba560-292b-4c13-91a7-0ab49e3a11d0
名前 / ファイル | ライセンス | アクション |
---|---|---|
MJI_26-1 (566.3 kB)
|
Item type | Journal Article(1) | |||||
---|---|---|---|---|---|---|
公開日 | 2017-06-23 | |||||
タイトル | ||||||
タイトル | TMEPAI genome editing in triple negative breast cancer cells | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源 | http://purl.org/coar/resource_type/c_6501 | |||||
タイプ | journal article | |||||
著者 |
Wardhani, Bantari W.K.
× Wardhani, Bantari W.K.× Puteri, Meidi U.× Watanabe, Yukihide× Louisa, Melva× Setiabudy, Rianto× Kato, Mitsuyasu |
|||||
著者別名 |
渡邊, 幸秀
× 渡邊, 幸秀× 加藤, 光保 |
|||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Background: Clustered regularly interspaced short palindromic repeats/CRISPR-associated 9 (CRISPR/Cas9) is a powerful genome editing technique. It consists of RNA-guided DNA endonuclease Cas9 and single guide RNA (gRNA). By combining their expressions, high efficiency cleavage of the target gene can be achieved, leading to the formation of DNA double-strand break (DSB) at the genomic locus of interest which will be repaired via NHEJ (non-homologous end joining) or HDR (homology-directed repair) and mediate DNA alteration. We aimed to apply the CRISPR/Cas9 technique to knock-out the transmembrane prostate androgen-induced protein (TMEPAI) gene in the triple negative breast cancer cell line. Methods: Designed gRNA which targets the TMEPAI gene was synthesized, annealed, and cloned into gRNA expression vector. It was co-transfected into the TNBC cell line using polyethylenimine (PEI) together with Cas9-GFP and puromycin resistant gene vector. At 24-hours post-transfection, cells were selected by puromycin for 3 days before they were cloned. Selected knock-out clones were subsequently checked on their protein levels by western blotting. Results: CRISPR/Cas9, a genome engineering technique successfully knocked-out TMEPAI in the Hs578T TNBC cell line. Sequencing shows a frameshift mutation in TMEPAI. Western blot shows the absence of TMEPAI band on Hs578T KO cells. Conclusion: TMEPAI gene was deleted in the TNBC cell line using the genomic editing technique CRISPR/Cas9. The deletion was confirmed by genome and protein analysis. |
|||||
書誌情報 |
Medical journal of Indonesia 巻 26, 号 1, p. 14-18, 発行日 2017-05 |
|||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 2252-8083 | |||||
書誌レコードID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AA1113872X | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | 10.13181/mji.v26i1.1871 | |||||
権利 | ||||||
権利情報 | Copyright @ 2017 Authors. | |||||
権利 | ||||||
権利情報 | This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original author and source are properly cited. | |||||
著者版フラグ | ||||||
値 | publisher | |||||
出版者 | ||||||
出版者 | University of Indonesia, Faculty of Medicine |