多重基因編輯於作物精準育種之應用

Abstract

多重基因編輯是新興精準育種的高效率工具之一，由於可以同時編輯基因組中2 個或多個不同的DNA位點，故適用於數量性狀基因或同源基因家族所調控重要性狀之探勘。操作多重基因編輯的系統可概略分成ZFNs、TALENs 及CRISPR/Cas 三種系統，其中以CRISPR/Cas 操作較為簡單、編輯效率較高以及具有成本較低的優勢，因此成為目前主流。本篇主要介紹以CRISPR/Cas 為基礎，進而發展出的多重基因編輯系統。文中另將針對編輯目的、載體構築表現系統、多gRNA 表現方式，以及在水稻、小麥、玉米、油菜、番茄及馬鈴薯等糧食與經濟作物的成功實例加以說明，冀能作為研究人員進行相關研究之參考依據。 Multiple gene editing is one of the highly efficient tools for novel precision breeding. Since two or more different DNA sites in the genome can be edited simultaneously, it is suitable for the exploration of important traits regulated by quantitative trait genes or homologous gene families. The systems of multiplex gene editing can be roughly divided into three modes: zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas. Among them, CRISPR/Cas has become the mainstream due to its simple operation, high editing efficiency, and low cost. Therefore, this article mainly discusses the advanced multiple gene editing system that was developed based on CRISPR/Cas. Furthermore, the subject will focus on the editing purpose, vector construction expression system, multiple gRNA expression methods, and successful examples in rice, wheat, corn, rape, tomato, potato and other food and economically important crops. We hope that this article can serve as a reference for researchers conducting related research.