CRISPR/Cas9 基因剔除快手

CRISPR/Cas9: A Powerful Tool for Genome Editing

基因編輯技術是一項幫助科學家了解基因功能的重要工具，可藉由基因剔除和基因轉殖來了解一個基因的表達對於生物體的影響，或作為人類疾病的模式。傳統的基因編輯技術是利用一系列耗時、耗力、效率低且繁瑣的同源互換技術來建構目標載體；相對於傳統技術，使用遺傳工程改造的核酸酶讓基因編輯技術更有效率，其中創新的CRISPR/Cas9 系統是由Cas9 蛋白和嚮導RNA 的結合來對DNA 進行修飾，具有設計簡單、製備過程容易的特性，大幅減少了基因編輯小鼠產製的時間。CRISPR/Cas9 系統具有高度應用性，未來將改寫科學家目前所使用的研究方法，以及能為人類疾病的研究帶來更多貢獻。

Genetic modification is the most critical technology in biology to investigate function of gene, which can be addressed via loss-of-function or gain-of-function mutations. Targeted genome editing in making knockout mice is traditionally achieved by sequential recombination in embryonic stem cell, and is time- and labor-consuming. The prokaryotic adaptive immunity mediated by clustered regularly interspaced short palindromic repeat (CRISPR) is an innovative approach for generating RNA-guided nucleases, such as Cas9. CRISPR/Cas9 is a powerful tool in introducing double strand break, which facilitates indel (insertion or deletion) mutation via non-homologous end joining (NHEJ) or precise incorporation of exogenous DNA fragments into target loci through homology-directed repair(HDR). Given the high-efficiency and broad-applicability of CRISPR/ Cas9, knockout and knockin mice can be generated in two months, and modified version of the CRISPR system can control specific gene expression in cells. Although the off-target and mosaicism issue of CRISPR/Cas9 systems remain to be fully uncovered, this simple-to-use and easy-to-design tool will transform and shed light on biomedical research.