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Patterson, A. T.

Publications and source records attributed to Patterson, A. T..

2 recordsLinked to original sources

Harnessing Plasmid Crosstalk in Transcription Factor Mediated Cell-Free Biosensors

Plasmid crosstalk--unexpected changes to protein expression levels due to interactions between genetic cassettes in cell-free systems--complicates the creation of multi-plasmid cell-free tools. While the potential underlying mechanisms for crosstalk have been previously investigated, the practical impact of crosstalk on the implementation of cell-free genetic circuits has not been thoroughly examined. Here, we contextualize plasmid crosstalk in genetic circuits by examining its impact on the design and performance of multiple, diverse transcription factor biosensors. Guided by a deeper understanding, we harness crosstalk to enhance cell-free biosensor performance. Together, these findings demonstrate that plasmid crosstalk can serve as a tunable property in the development of cell-free genetic circuits.

synthetic biology↗

A modular cell-free protein biosensor platform using split T7 RNA polymerase

Conventional laboratory protein detection techniques are not suitable for point-of-care (POC) use because they require expensive equipment and laborious protocols, and existing POC assays suffer from long development timescales. Here, we describe a modular cell-free biosensing platform for generalizable protein detection that we call TLISA (T7 RNA polymerase-Linked ImmunoSensing Assay), designed for extreme flexibility and equipment-free use. TLISA uses a split T7 RNA polymerase fused to affinity domains against a protein. The target antigen drives polymerase reassembly, inducing reporter expression. We characterize the platform, then demonstrate its modularity by using 16 affinity domains against four different antigens with minimal protocol optimization. We show TLISA is suitable for POC use by sensing human biomarkers in serum and saliva with a colorimetric readout within one hour and by demonstrating functionality after lyophilization. Altogether, this technology could have potentially revolutionary impacts, enabling truly rapid, reconfigurable, equipment-free detection of virtually any protein.

synthetic biology↗