Development of Infertility Technology in Transgenic Fluorescent Ornamental Fish (I)

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Project title

Code/計畫編號

MOST103-2622-B019-001-CC1

Translated Name/計畫中文名

基因轉殖螢光觀賞魚之不孕技術開發( I )

Project Coordinator/計畫主持人

Hong-Yi Gong

Funding Organization/主管機關

National Science and Technology Council

Co-Investigator(s)/共同執行人

胡紹揚
吳金洌

Department/Unit

Department of Aquaculture

Website

https://www.grb.gov.tw/search/planDetail?id=8316916

Year

2014

Start date/計畫起

01-06-2014

Expected Completion/計畫迄

31-05-2015

Bugetid/研究經費

1500千元

ResearchField/研究領域

生物技術(農)
漁業

" 觀賞魚產業是精緻的養殖產業，為台灣農業生技發展重點項目之ㄧ。台灣在 2010年 6 月由芝林公司與中研院合作開發出全球首例之中型螢光轉基因神仙魚與九間波羅，因此在全球基因轉殖觀賞魚產業上具有技術領導地位。中型螢光神仙魚(Pterophyllum scalare) 與九間波羅 (Amatitlania nigrofasciata) 均屬於中型慈鯛科魚類，因獨創性與新穎性高，預估通過生物安全評估後屆時國外市場將達 1 千 6 百萬美元產值。由此可知，台灣基因轉殖螢光觀賞魚在全球觀賞魚產業已奠定成熟基礎，並且已展現可預期的經濟價值，具有潛在龐大經濟商機。然而因全球目前對基因轉殖生物安全具有疑慮，憂心一旦基因轉殖魚類流放至野外時，將導致生態平衡的破壞與基因流佈(gene flow)等問題，因此造成基因轉殖觀賞魚行銷受限，而觀賞魚業者也擔心一旦基因轉殖觀賞魚遭人私下繁殖，將造成價格低落導致行銷的困難，因此開發魚類不孕控制技術為拓展觀賞魚產業發展的核心技術。基因轉殖魚類的不孕控制研究是一個重要的生物安全性考量因子，且須為可誘導性方能將基因轉殖魚類傳承下來。本計畫將選殖神仙魚與九間波羅性腺專一性抗穆勒氏管賀爾蒙(Anti-Müllerian hormone; amh)基因與啟動子，探討慈鯛科 amh 基因對性腺分化與成熟之影響，並利用生殖細胞專一性 amh 啟動子表現誘發性毒性蛋白，藉由基質的浸泡或光照處理來誘發生殖細胞死亡產生不具生殖能力之基因轉殖神仙魚與九間波羅。本計劃針對神仙魚與九間波羅為目標，開發慈鯛科誘導性不孕控制技術，預期將完成生殖弱化之螢光轉基因觀賞神仙魚與九間波羅，協助儘早通過基因轉殖田間隔離試驗生物安全性評估，完成基因轉殖觀賞神仙魚與九間波羅上市行銷之目標。本計劃擬利用 Piwi基因表現在初始生殖細胞(PGC)及雌雄生殖細胞精細胞與卵細胞發育過程，而 AMH基因表現在生殖細胞周圍的體細胞精巢之 Sertoli細胞與卵巢之濾泡細胞。擬選殖魚類性腺專一性 Piwi與 AMH基因啟動子表現 NTR誘導性毒性蛋白，藉由處理基質 Mtz 分別破壞初始生殖細胞及性腺之生殖細胞與周圍的體細胞 Sertoli 細胞或濾泡細胞以達到完全不孕之目標。此計畫經由 TALEN或 CASPRE-Cas技術將神仙魚 Piwi1, Piwi2及 AMH基因剔除，可探討慈鯛科 Piwi 及 AMH基因對性腺分化與成熟之影響，更有助於基因轉殖螢光斑馬魚及神仙魚通過生物安全評估系統，讓台灣人研發的創新與技術領先的基因轉殖中型螢光觀賞魚早日進入國際市場。 " "Ornamental fish industry is a fine aquaculture industry. It is also one of priority subjects of agriculture biotechnology in Taiwan. Since 2010, the first transgenic fluorescent angle fish (Pterophyllum scalare) and cichlid (Amatitlania nigrofasciata) were established by Academia Sinica and Jy Lin Company in Taiwan, thus Taiwan possess the technical leadership in the industry of transgenic ornamental fish. Transgenic fluorescent angle fish and cichlid belong to medium-sized cichlid with originality and novelty. It was expected that the market will reach 16 million while obtaining approval of biosafety assessment. From the fact we know that transgenic fluorescent ornamental fish in Taiwan has been established a mature foundation in the global ornamental fish industry, and demonstrated potentially with a hung economic value. However, because of the international concern in genetically modified (GM) fish which may cause the threat of extinction and gene-flow in the wild, the global sale of transgenic fish is restricted; moreover the impediment of sale for ornamental fish is worry about the private breeding, which will cause the price fall down. For these reasons, the development of infertile control technology is considered to be core technology for ornamental fish industry requirements. The technology of infertile control for transgenic fish is considered to be prominent issue for meet the GMO biosafety and this technology must be inducible; otherwise the transgenic fish can’t be inherited. In this project, we will use the expression specificity of Piwi and AMH promoters from zebrafish, tilapia or angelfish to express an inducible toxin protein nitroreductase in the primordial germ cells, different developmental stages of germ cells including spermatogonia and oogonia (by Piwi promoter) and gonad surrounding somatic cells including Sertoli cells and follicle cells (by AMH promoter). The cytotoxin will destroy germ cells and surrounding somatic cells of gonads in both male and female transgenic fish and results in infertility by the treatment of Mtz substrate solution. In addition, we will knockout Piwi1, Piwi2 and AMH genes in angelfish by using TALEN or CASPRE-Cas techniques to study the functions of Piwi and AMH genes in gonad differentiation and maturation of cichlid fish. We expect to perform the infertility of transgenic fluorescent zebrafish and angelfish to achieve approval of biosafety assessment and facilitate the middle-sized transgenic fluorescent ornamental fish created by innovation and leading techniques of Taiwaneses swimming into the international markets in the nearby future. "

Keyword(s)

基因轉殖觀賞魚
不孕控制
Piwi
抗穆勒氏賀爾蒙
transgenic ornamental fish
infertility control
AMH

DSpace CRIS

Development of Infertility Technology in Transgenic Fluorescent Ornamental Fish (I)

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