Aptazyme-mediated direct modulation of post-transcriptional sgRNA level for conditional genome editing and gene expression

作者：Chen, Hao; Li, Yanqing; Du, Chunhua; Li, Yan; Zhao, Junli; Zheng, Xiaojing; Mao, Qinwen; Xia, Haibin^*

来源：Journal of Biotechnology, 2018, 288: 23-29.

DOI：10.1016/j.jbiotec.2018.10.011

• Genome editing
• Cas9
• sgRNA
• Theophylline
• Aptazyme
• Off-target effect

RNA-guided endonuclease Cas9 derived from microbial CRISPR-Cas adaptive immune systems is a powerful tool for genome editing, which has been widely used in eukaryotic systems, prokaryotic systems, and plants. However, the off-target effects caused by Cas9/sgRNA remain a major concern. Currently, the efforts to reduce the off-target effects mainly focus on improving the targeting specificity of sgRNA/Cas9, regulating the activity of the Cas9 protein or the sgRNA, and controlling the time window of their expression. In this study, a novel system was established to regulate the post-transcriptional sgRNA level by small molecule-controlled aptazyme. This system was shown to reduce the off-target effects caused by Cas9/sgRNA, while enabling precise temporal control over gene editing and regulatory activity. This new system could provide a potentially safer and more powerful tool for genome editing and therapeutic application.

• 出版日期2018-12-20
• 单位陕西师范大学; 西北大学