Detection of MicroRNAs Using Synthetic Toehold Switch in Mammalian Cells

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2774; p. 243
• Main Authors: Zhao, Yuwen, Poudel, Pratima, Wang, Shue
• Format: Journal Article
• Language: English
• Published: United States 2024
• Subjects: miRNA-based switch; Synthetic biology; MicroRNA; Synthetic toehold switch
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-3718-0_16

Engineering synthetic gene circuits to control cellular functions has a broad application in the field of synthetic biology. Synthetic RNA-based switches that can operate at the transcriptional and posttranscriptional level have also drawn significant interest for the application of next-generation therapeutics and diagnostics. Thus, RNA-based switchable platforms are needed to report dynamic cellular mechanisms which play an important role in cell development and diseases. Recently, several RNA-based switches have been designed and utilized for biosensing and molecular diagnostics. However, miRNA-based switches have not been well established or characterized, especially for eukaryotic translational control. Here, we designed a novel synthetic toehold switch for detection of exogenously and endogenously expressed miRNAs in CHO, HeLa, HEK 293, and MDA-MB-231 breast cancer cells. Multiplex detection of miR-155 and miR-21 was tested using two toehold switches to evaluate the orthogonality and programmability of this synthetic platform.