Hao Huanhuan, Wang Xiaofei, Jia Haiyan, Yu Miao, Zhang Xiaoyu, Tang Hui, Zhang Liping
Key Laboratory of Microbial Diversity Research and Application of Hebei Province, Key Discipline of Biological Engineering of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
Key Laboratory of Microbial Diversity Research and Application of Hebei Province, Key Discipline of Biological Engineering of Hebei Province, College of Life Sciences, Hebei University, Baoding, 071002, China.
Anal Biochem. 2016 Sep 15;509:118-123. doi: 10.1016/j.ab.2016.07.008. Epub 2016 Jul 9.
Large chromosomal modifications have been performed in natural and laboratory evolution studies and hold tremendous potential for use in foundational research, medicine, and biotechnology applications. Recently, the type II bacterial Clustered Regularly Interspaced Short Palindromic Repeat and CRISPR-associated (CRISPR/Cas9) system has emerged as a powerful tool for genome editing in various organisms. In this study, we applied the CRISPR/Cas9 system to preform large fragment deletions in Saccharomyces cerevisiae and compared the performance activity to that of a traditional method that uses the Latour system. Here we report in S. Cerevisiae the CRIPR/Cas9 system has been used to delete fragments exceeding 30 kb. The use of the CRISPR/Cas9 system for generating chromosomal segment excision showed some potential advantages over the Latour system. All the results indicated that CRISPR/Cas9 system was a rapid, efficient, low-cost, and versatile method for genome editing and that it can be applied in further studies in the fields of biology, agriculture, and medicine.
在自然进化和实验室进化研究中已经进行了大规模染色体修饰，其在基础研究、医学和生物技术应用方面具有巨大的潜力。最近，II型细菌成簇规律间隔短回文重复序列及其相关蛋白（CRISPR/Cas9）系统已成为一种用于各种生物体基因组编辑的强大工具。在本研究中，我们应用CRISPR/Cas9系统在酿酒酵母中进行大片段缺失，并将其性能活性与使用拉图尔系统的传统方法进行比较。在此我们报告，在酿酒酵母中，CRIPR/Cas9系统已被用于删除超过30 kb的片段。使用CRISPR/Cas9系统进行染色体片段切除显示出比拉图尔系统更具潜在优势。所有结果表明，CRISPR/Cas9系统是一种快速、高效、低成本且通用的基因组编辑方法，可应用于生物学、农业和医学领域的进一步研究。
