Ni Peiling, Zhang Qian, Chen Haixia, Chen Lingyi
State Key Laboratory of Medicinal Chemical Biology, 2011 Collaborative Innovation Center of Tianjin for Medical Epigenetics, College of Life Sciences, Nankai University, Tianjin, China.
Biotechniques. 2014 Apr 1;56(4):198-201. doi: 10.2144/000114160. eCollection 2014.
Removing an antibiotic resistance gene allows the same antibiotic to be re-used in the next round of genetic manipulation. Here we applied the CRISPR/Cas system to disrupt the puromycin resistance gene in an engineered mouse embryonic stem cell line and then re-used puromycin selection in the resulting cells to establish stable reporter cell lines. With the CRISPR/Cas system, pre-engineered sequences, such as loxP or FRT, are not required. Thus, this technique can be used to disrupt antibiotic resistance genes that cannot be removed by the Cre-loxP and Flp-FRT systems.
去除抗生素抗性基因可使同一种抗生素在新一轮基因操作中再次使用。在此，我们应用CRISPR/Cas系统破坏一种工程化小鼠胚胎干细胞系中的嘌呤霉素抗性基因，然后在所得细胞中重新使用嘌呤霉素筛选以建立稳定的报告细胞系。使用CRISPR/Cas系统时，不需要预先设计的序列，如loxP或FRT。因此，该技术可用于破坏无法通过Cre-loxP和Flp-FRT系统去除的抗生素抗性基因。
