Abstract: Provided herein, in one aspect, is a composition for in vitro transcription and translation, comprising: a treated cell lysate derived from one or more host cells such as bacteria, archaea, plant or animal; a plurality of supplements for gene transcription and translation; an energy recycling system for providing and recycling adenosine triphosphate (ATP); and one or more exogenous additives. Methods for making and using the same are also provided.

Abstract: Provided herein, in one aspect, is a composition for in vitro transcription and translation, comprising: a treated cell lysate derived from one or more organisms such as bacteria, archaea, plant or animal; a plurality of supplements for gene expression; an energy recycling system for providing adenosine triphosphate and recycling adenosine diphosphate; and an engineered propeptide operably linked to a stabilizing domain. Methods for making and using the same are also provided.

Abstract: Provided herein are methods and compositions for screening using an in vitro transcription/translation (TXTL) system. In some embodiments, a screening method can include: providing a library of genetic candidates to be screened, each genetic candidate encoding one or more protein of interest; expressing each genetic candidate in an in vitro transcription/translation (TXTL) system, thereby producing a plurality of compositions each comprising the one or more protein of interest; and subjecting the plurality of compositions to an assay for assessing a desired phenotype, thereby determining whether the one or more protein of interest exhibits the desired phenotype.

Abstract: The present disclosure relates to cell-free compositions and methods for making and using the same. In one aspect, the composition includes: an extract derived from one or more organisms; one or more proteins of interest, wherein the one or more proteins are expressed from one or more nucleic acids exogenous to the extract and/or by the one or more organisms, wherein preferably the one or more proteins react with a substrate to produce a product; and one or more O2, O—, or H2O2 scavengers. The composition may be oxygen-deprived. The composition may also include an energy recycling system.

Abstract: Compositions and methods disclosed herein relate to improved in vitro cell-free systems for various biological reactions. In one aspect, a composition of the present disclosure includes: a cell-free extract derived from an organism; a nucleic acid; and an organelle that is heterologous to the organism.

Abstract: Provided herein are methods and arrangements and related cell-free biomolecular breadboards configured to design, build, implement, debug, and/or test a genetic circuit to be operated in a target environment, by testing in a cell-free system under conditions of the target environment, molecular components of the genetic circuit and/or combinations thereof to select the molecular components and/or combinations thereof of a genetic circuit operative in the target environment.

Abstract: Provided herein, in one aspect, is a composition for in vitro transcription and translation, comprising: a treated cell lysate derived from one or more host cells such as bacteria, archaea, plant or animal; a plurality of supplements for gene transcription and translation; an energy recycling system for providing and recycling adenosine triphosphate (ATP); and one or more exogenous additives. Methods for making and using the same are also provided.

Abstract: Provided herein, in one aspect, is a composition for in vitro transcription and translation, comprising: a treated cell lysate derived from one or more organisms such as bacteria, archaea, plant or animal; a plurality of supplements for gene expression; an energy recycling system for providing adenosine triphosphate and recycling adenosine diphosphate; and an engineered propeptide operably linked to a stabilizing domain. Methods for making and using the same are also provided.

Abstract: Provided herein are methods and compositions for rapid assembly of genetic modules, as well as seamless transition from in vitro to in vivo testing of genetic constructs.

Abstract: Provided herein are methods and compositions for rapid assembly of genetic modules, as well as seamless transition from in vitro to in vivo testing of genetic constructs.

Abstract: Provided herein are methods and arrangements and related cell-free biomolecular breadboards configured to design, build, implement, debug, and/or test a genetic circuit to be operated in a target environment, by testing in a cell-free system under conditions of the target environment, molecular components of the genetic circuit and/or combinations thereof to select the molecular components and/or combinations thereof of a genetic circuit operative in the target environment.