(2aq) Nucleic Acid Detection By Target-Assisted Synthesis of Enzyme Reporters in a Cell-Free Protein Synthesis System

Sensitive and accurate detection of nucleic acids is a crucial element for early diagnostics and effective management of diseases. Most of the present methods for nucleic acid detection are based on the amplification of target sequences by PCR and related method However, these methods require specialized equipment and reagents. In this study, we present a novel approach that involves the measurement of nucleic acids by the biological activity of enzymes, which are produced in response to the target nucleic acid sequences. In this method, dubbed target-assisted synthesis of reporters (TASR), a sensor DNA recognizes the target sequence and activates the process of cell-free protein synthesis. Through the cascade of transcription and translation reactions, the recognized target sequence is converted into amplified number of functional reporter proteins. Taking advantage of the modular nature of cell-free protein synthesis, the TASR assay readily converts target sequences to different reporter proteins, which greatly improves the flexibility of nucleic acid detection without the use of complicated devices and laboratory settings.