2013-05-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/646726摘要：研究人類所有的基因的功能和疾病的機制是醫學界最重要的工作之一，而小鼠無疑地 是最能模擬人類生理機制的模型動物之一。因此基因突變小鼠成為研究人類基因功能 與疾病機制最重要的工具。在 2006 年，歐洲及加拿大宣佈將在胚胎幹細胞進行 “conditional knockout”，同年，美國 NIH 支持 KOMP (Knockout Mouse Project)將再增 加一萬個基因剔除鼠，預計國際上總花費近一億美金。來自國際的訊息顯示現今的基 因剔除技術只能完成 60%的人類基因剔除，故須發展新的 knockout 技術，但最後仍約 有 10-20%基因須要逐一進行剔除。總體而言，所有的基因剔除胚胎幹細胞，都需要利 用幾種技術再結合胚體，才能產製出剔除小鼠，因此須要更多基因轉殖核心實驗室來 協助科學家加速完成基因功能的分析。有鑑於此，本團隊的宗旨之一就是建立基因剔 除或置入小鼠核心設施，提供技術協助研究者製造基因剔除或置入小鼠。自 2005 年主 持人承接 NRPGM 的基因轉殖鼠核心設施(A4)計畫以來，已接受來自 88 個研究單位之 146 位研究學者申請的 301 個基因剔除計畫。本核心至今已協助使用者產出超過 203 種可繼代遺傳的基因剔除或置入小鼠。主持人認為計畫的順利推行主要因為本核心主 動與使用者討論出基因剔除或置入的策略。此外，利用 Recombineering 方法可以很容 易將 loxP 與 FRT 序列加入基因剔除載體，使得一個載體就能同時作 conditional 及 conventional 剔除，免除胎死的遺憾。另一個是提供大規模 Southern blotting 的服務， 使 各個服務案件得以順利進行。基於此，主持人提出本計畫希望繼續已有的基礎與成果， 目標在於 1.繼續提供服務，2.研發相關科技，3. 深入與使用者的合作研究計畫，與 4. 推廣核心設施等工作，幫助國內學者完成基因剔除或置入小鼠，推動其研究更上層樓。 主持人認為本團對目前已成為最能提供基因剔除鼠技術給在台灣從事生物醫學研究者 的團隊之一，主持人期望本團隊經由此計畫，一方面能強化本院基因轉殖鼠核心設施， 最終目標是能成為國內基因體醫學研究的重要支援單位，強化國內利用基因剔除或置 入小鼠研究人類基因功能與疾病的研究水準。<br> Abstract: The biomedical sciences community has committed to expanding our understanding of gene function and various mechanisms involved in the development of human diseases. Mouse has been one of the best model organisms for studying human gene functions. In 2006, Europe and Canada announced together to produce more than 30,000 knockouts. The same summer, the U.S. NIH announced the Knockout Mouse Project (KOMP), which would add another 10,000 to the list. There is an urgent need for funding more transgenic mouse core facilities to not only increase the 60% coverage of the entire genome (targeting efficiency) the international efforts can reach to 100%, but to verify the targeted ES cell clones generated by these international cores and converting these clones into to live mice. Moreover, there is still overall a 10-20% of the genome that need to be targeted individually. The Principle investigator has trained and organized a team capable of performing knockout technology. The P.I.’s team has been in charge of the (A4) Transgenic Mouse Models core (TMMC) under the support of the NRPGM (National Research Program for Genomic Medicine) since the fiscal year 2005. We have received nearly 301 knockout and/or knock-in projects from 146 P.I. over 88 research institutes in this nation since granted. This core has helped our users to generate over 203 germline-transmitted knockout (KO) mice. We think that our success are mainly due to two strategies: one is providing free consultation on targeting strategy to users and providing the service for targeting construct production using the “recombineering method” to shorten the time required to make a construct using traditional cloning strategy, which is very attractive to the users who are not familiar with but would like to try using knockout/knock-in mice in their research. In addition to that, the “recombineering method” can easily introduce loxP and FRT sites in the vector so that one single vector can server both conventional and conditional KO purposes, which can solve the embryonic lethality problems. The other is providing the service for Southern blotting screening for targeted ES clones. We submit this proposal to continue on providing our expertise on ES cell gene targeting as well as the following goals, 1. Service, 2. Research and development (R&D), 3. Collaborative research and 4. Disseminations. I think the team is in its best position to serve the academic community. The ultimate objective of this project is to facilitate the use of mice in this National as models for studying gene functions and phenotypes of human diseases.