METHOD FOR PERFORMING GENETIC MODIFICATION UNDER A DRUG-FREE ENVIRONMENT AND COMPONENTS THEREOF

專利類型

發明

專利國別 (專利申請國家)

美國

專利申請案號

12/588,708

專利證號

US 9,790,489 B2

專利獲證名稱

METHOD FOR PERFORMING GENETIC MODIFICATION UNDER A DRUG-FREE ENVIRONMENT AND COMPONENTS THEREOF

專利所屬機關 (申請機關)

長庚大學

獲證日期

2018/10/17

技術說明

本專利申請案描述一已發展成功無需以药物來篩選之遺傳エ程技術平臺。本創新的遺傳エ程技術，除了可運用於哺乳動物及其細胞株的基因體改造外，並針對現今技術無法克服處於發育及分化過程中不表達或被抑止表達之基因的操作上，提供了有效的解決方法。本專利亦提供了具體且具高效率的操作步驟，以方便獲取剔除幹細胞中不表達基因之幹細胞株，其內容包括：(a)建立剔除未表達基因之鼠胚胎幹細胞文庫；(b)系統化地評估在基因剔除文庫中捕捉到之未表達基因的比例；(c)建立双報導基因系统來增加微量表達基因的捕捉。本創新方法有別於目前遺傳エ程的技術於：(1)減少相同基因剔除的細胞株重覆地出現於文庫中；(2)有效的剔除維持幹細胞多元分化能力時所需被抑止表達的基因；(3)免除基因轉殖時的药物篩選過程。同時，也避免抗生素基因留存於基因改造動物(transgenic animal)之基因體中，而間接造成環境危害。 A drug-free selection method for the technology of genetic engineering is described. The present invention relates to genetically engineering vertebrates including mammals and mammalian cells, with a especial focus in the method of targeting silent or repressed genes in a cell line and/or mouse stem cell. The advantages of using the present invention include: (1) minimizing the bias of targeting genes located at “hot spots”, the drawbacks seen in all of gene targeting technologies currently available; (2) advantageously targeting key genes involving in critical developmental decisions but are silenced or repressed in embryonic or other type of stem cells; and (3) Avoiding the need of introducing antibiotic genes into the host genome and in turn eliminating potential threats of drifting antibiotic resistant genes into environment.

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