Zhang Wenhui, Zhao Shi, Wang Mengyuan, Lou Chunbo, Xiang Yanhui, Wu Qiong
Center for Cell and Gene Circuit Design, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China.
State Key Laboratory of Green Biomanufacturing, MOE Key Lab. Bioinformatics, Center for Synthetic and Systems Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Synth Syst Biotechnol. 2025 Apr 8;10(3):827-834. doi: 10.1016/j.synbio.2025.03.011. eCollection 2025 Sep.
Synthetic RNA-based switches provide distinctive merits in modulating gene expression. Simple and flexible RNA-based switches are crucial for advancing the field of gene regulation, paving the way for innovative tools that can sense and manipulate cellular processes. In this research, we have developed programmable ribozymes that are capable of suppressing gene expression in response to specific, endogenously expressed trigger RNAs. We engineer ribozymes by introducing upstream antisense sequences (anti-ribozymes) to inhibit the self-cleaving activity of the hammerhead ribozyme and open the expression of the target gene. The trigger RNA is designed to recognize and bind to complementary sequences within the anti-ribozymes, thereby inhibiting their ability to direct protein synthesis. The anti-ribozyme performance is optimized by regulating the essential sequence modules that play a crucial role in determining the specificity and efficiency of the anti-ribozyme's interaction with its trigger RNA. By applying this switch mechanism to various ribozyme designs, we have shown that it is possible to achieve control over gene expression across a wide range of trigger RNAs. By exploiting these programmable anti-ribozymes, we aim to create a powerful tool for controlling gene expression in mammalian cells, which could have important implications for basic research, disease diagnosis, and therapeutic interventions.
基于合成RNA的开关在调控基因表达方面具有独特优势。简单且灵活的基于RNA的开关对于推动基因调控领域的发展至关重要，为能够感知和操纵细胞过程的创新工具铺平了道路。在本研究中，我们开发了可编程核酶，其能够响应特定的内源性表达触发RNA来抑制基因表达。我们通过引入上游反义序列（抗核酶）来设计核酶，以抑制锤头状核酶的自我切割活性并开启靶基因的表达。触发RNA被设计用于识别并结合抗核酶内的互补序列，从而抑制其指导蛋白质合成的能力。通过调节在决定抗核酶与其触发RNA相互作用的特异性和效率方面起关键作用的基本序列模块，优化抗核酶性能。通过将这种开关机制应用于各种核酶设计，我们已表明能够在广泛的触发RNA范围内实现对基因表达的控制。通过利用这些可编程抗核酶，我们旨在创建一种用于控制哺乳动物细胞中基因表达的强大工具，这可能对基础研究、疾病诊断和治疗干预具有重要意义。
