以甜瓜原生質體短暫性表現轉殖系統快速評估基因編輯效果

常間回文重複序列叢集/Cas9 蛋白質系統 (clustered regularly interspaced short palindromic repeats/Cas9; CRISPR/Cas9) 為近年來興起之基因組編輯技術，亦是目前基因功能研究及精準育種課題的重要新興核心工具。對比現行農桿菌法媒介轉形過程所需時間冗長，本研究旨建立基因組編輯系統的快速導入平台，以利進行基因編輯試驗的先期評估。首先，本研究選用葫蘆科之甜瓜 (Cucumis melo L.) 作為主要材料，建立葉片原生質體短暫性表現系統平台，其轉殖效率約可達20%。同時，利用甜瓜真核轉譯起始因子4E (eukaryotic translation initiation factors 4E; Cm-eIF4E) 作為標的基因，輔以葉片原生質體短暫性表現系統，進行甜瓜細胞之基因組編輯效應評估。 The clustered regularly interspaced short palindromic repeats/Cas9 (CRISPR/Cas9) system is the most popular strategy for genomic editing technology in recent years, and has been a powerful tool for gene function verification and precision plant breeding. To replace the current time-consuming Agrobacterium-mediated method, this study aims to establish a rapid transient expression platform for the preliminary evaluation of genomic editing system. First of all, leaf protoplasts of Cucumis melo L. were used to establish the transient gene expression system, and the best transformation efficiency reached approximately 20%. Simultaneously, the melon eukaryotic translation initiation factors 4E (eIF4E) was used as a target gene to evaluate the genome editing efficiency in the transient expression system.

Subjects

原生質體

基因組編輯

嚮導RNA

甜瓜真核轉譯起始因子4E

Protoplast

Genomic editing

Dual gRNA

Cm-eIF4E

Publisher

農業試驗所

Type

journal article

File(s)

Name

70-3-5.pdf

Size

2.56 MB

Format

Adobe PDF

Checksum

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