Abstract: Disclosed herein are biosensors, which may be made from cell lysates, purified enzymes, or a combination thereof, for testing for the presence of a pathogen, such as SARS-CoV-2. The biosensor may be used in the context of a paper-based test, such as a colorimetric COVID-19 test. Methods of using the biosensor are also disclosed.

Abstract: Disclosed herein are biosensors including transcription, translation, and coupled transcription/translation systems. The biosensors may be made from cell lysates, purified enzymes, or a combination thereof. The biosensors may include an inhibitor, and may include a reporter. The biosensor may be supplied in a kit for testing for a disease or condition. Methods of using the biosensor are also disclosed.

Abstract: The present disclosure as disclosed in various embodiments is related to a bioassay for rapid screening and detection of bioactive ligands and screening for potential pharmaceuticals. In various embodiments are disclosed cell-free methods, systems, and kits for detecting bioactive ligands such as endocrine disrupting chemicals or nuclear hormone receptor modulators in a biological or environmental sample or for screening compounds for potential pharmaceutical application such as potential nuclear hormone receptor modulator pharmaceuticals.

Abstract: The present disclosure provides methods for incorporating at least one non-natural amino acid into a polypeptide using a cell-free protein synthesis which includes a cell-free extract and is deficient in endogenous tRNA. The methods include providing the synthesis system with at least one non-natural amino acid, at least one orthogonal tRNA and one orthogonal aminoacyl-tRNA synthetase which aminoacylates the corresponding orthogonal tRNA with the non-natural amino acid. The methods may also include removing the endogenous tRNA in the cell-free protein synthesis system by treating the synthesis system with a ribonuclease to degrade the endogenous tRNA and providing the synthesis system with a minimal set of tRNAs for one or more natural amino acids and the corresponding amino-acyl tRNA synthetases, such that the lack of some tRNA will enable unique codons to encode for non-natural amino acid incorporation with no or a minimal amount of completion from the tRNA present.

Abstract: Methods are provided for the utilization of bacterial cell-free extracts in the synthesis of high yields of virus like particles with encapsidated cargo.

Abstract: Compositions and methods are provided for the control of direct protein attachment to virus like particles where virus structural proteins that have been modified to comprise an unnatural amino acid at a pre-determined site are reacted with one or more “display” polypeptides that also comprise an unnatural amino acid at a pre-determined site in a one step reaction. The compositions of the invention are useful for various purposes where it is desirable to efficiently and directly attach multiple polypeptides to a single carrier entity, particularly where two or more different polypeptides are attached to a single carrier.

Abstract: Methods are provided for the utilization of bacterial cell-free extracts in the synthesis of high yields of virus like particles with encapsidated cargo.

Abstract: Methods are provided for the utilization of bacterial cell-free extracts in the synthesis of high yields of virus like particles.