Liu Yifan, Ge Huanhuan, Marchisio Mario Andrea
School of Pharmaceutical Science and Engineering, Tianjin University, Tianjin, China.
Front Bioeng Biotechnol. 2023 Sep 12;11:1267174. doi: 10.3389/fbioe.2023.1267174. eCollection 2023.
Among CRISPR-Cas systems, type V CRISPR-Cas12c is of significant interest because Cas12c recognizes a very simple PAM (TN) and has the ability to silence gene expression without cleaving the DNA. We studied how new transcription factors for the yeast can be built on Cas12c. We found that, upon fusion to a strong activation domain, Cas12c is an efficient activator. Its functionality was proved as a component of hybrid Boolean gates, i.e., logic circuits that mix transcriptional and translational control (the latter reached via tetracycline-responsive riboswitches). Moreover, Cas12c activity can be strongly inhibited by the anti-CRISPR AcrVA1 protein. Thus, Cas12c has the potential to be a new tool to control the activation of gene expression within yeast synthetic gene circuits.
在CRISPR-Cas系统中，V型CRISPR-Cas12c备受关注，因为Cas12c识别非常简单的PAM（TN），并且具有在不切割DNA的情况下使基因表达沉默的能力。我们研究了如何基于Cas12c构建用于酵母的新型转录因子。我们发现，与强激活结构域融合后，Cas12c是一种高效的激活剂。其功能作为混合布尔门（即混合转录和翻译控制的逻辑电路，后者通过四环素响应核糖开关实现）的一个组件得到了证明。此外，Cas12c的活性可被抗CRISPR蛋白AcrVA1强烈抑制。因此，Cas12c有潜力成为控制酵母合成基因电路中基因表达激活的新工具。
