基因編輯系統的發展暨其在作物育種上的應用

Gene Editing and Its Applications in Crop Breeding

王怡雯(Yi-Wen Wang) ； 謝昆廷(Kun-Ting Hsieh) ； 游舜期(Shuen-Chi You) ； 林大鈞(Da-Gin Lin) ； 陳良築(Liang-Jwu Chen)

《植物種苗》 18卷2期 (2016/06) Pp. 1-25

摘要

CRISPR／Cas9系統（Clustered regularly interspaced short palindromic repeats／CRISPR-associated protein）是一種新的基因編輯技術，此系統操作簡單，編輯效率高和成本低，已被廣泛應用，因此迅速取代ZFNs（Zinc finger nucleases）及TALENs（Transcription activator-like effector nucleases），成為新一代的基因組編輯技術。目前在植物中針對不同的物種，已經開發出具特異性的CRISPR／Cas9轉殖載體，而這些載體也已經成功應用於如水稻、玉米、小麥、大豆等作物之基因編輯。透過基因編輯不但可有效研究植物基因之功能，也有機會改良目標作物，增加其對生物性和非生物性逆境的抗性或是增加產量等，創造出良好且有利用價值的新品種。本文將介紹植物基因編輯之發展、CRISPR／Cas9發展歷史及原理、CRISPR／Cas9基因編輯在植物上之應用優勢以及操作步驟：包括sgRNA的設計、啟動子的選擇、如何篩選及檢測基因編輯是否成功等。最後，介紹基因編輯技術應用在作物育種上的實例與現況。

關鍵字

CRISPR／Cas9系統 ； 基因編輯 ； 作物育種

並列摘要

The CRISPR/Cas9 system (Clustered regularly interspaced short palindromic repeats/CRISPR-associated protein) is a novel gene editing technology. This technology is easy to operate, has high editing efficiency as well as low costs and has replaced ZFNs (Zinc finger nucleases) and TALENs (Transcription activator-like effector nucleases) methods to become the most popular gene editing technology. At present, specific CRISPR/Cas9 vectors have been developed and applied successfully in gene editing for rice, corn, wheat and soybean. In addition to study the function of genes, this technology also be used to create crops with novel and useful characteristics for increasing resistance to biotic- or abiotic- stresses and to enhance crop yields by editing the relevant key genes. This article will introduce the development of gene editing and the CRISPR/Cas9 system, its advantages and the CRISPR/Cas9 associated protocols when editing plant genes such as the design of sgRNA, the selection of promoters, and the screening and detection of edited gene sequences. Finally, examples and current progress of gene editing technology used in crop breeding are discussed.

並列關鍵字

CRISPR/Cas9 ； genome editing ； crop breeding