Improved production of recombinant proteins in E. coli, reliant on tightly controlled autoinduction, triggered by phosphate depletion in stationary phase. The process also provides an optimized autoinduction media, enabling routine batch production at various culture volumes where cells densities routinely reach ˜5-7 g cell dry weight per liter and offer protein titers above 2 g/L. The methodology has been validated with a set of diverse heterologous proteins and is of general use for the facile optimization of routine protein expression from high throughput screens to fed-batch fermentation.

Microbial consortia exert great influence over the physiology of humans, animals, plants, and ecosystems. However, difficulty in controlling their composition and population dynamics have limited their application in medicine, agriculture, biotechnology, and the environment. The approach disclosed herein provides an effective method to dynamically control population compositions in microbial consortia, which we demonstrate in the context of co-culture fermentations for chemical production. Co-culture fermentations can improve chemical production from complex biosynthetic pathways over monocultures by distributing enzymes across multiple strains, thereby reducing metabolic burden, overcoming endogenous regulatory mechanisms, or exploiting natural traits of different microbial species. However, stabilizing and optimizing microbial sub-populations for maximal chemical production remains a major obstacle in the field. An optogenetic circuit, called OptoTA, is disclosed for regulating a toxin-antitoxin system, which enables tunability of, e.g., Escherichia coli growth using only blue light. With the disclosed system, one can control population ratios of co-cultures of, e.g., E. coli and Saccharomyces cerevisiae containing different metabolic modules of biosynthetic pathways. Results reveal that intermediate light duty cycles improve chemical production by establishing optimal co-culture populations.

Microbial consortia exert great influence over the physiology of humans, animals, plants, and ecosystems. However, difficulty in controlling their composition and population dynamics have limited their application in medicine, agriculture, biotechnology, and the environment. The approach disclosed herein provides an effective method to dynamically control population compositions in microbial consortia, which we demonstrate in the context of co-culture fermentations for chemical production. Co-culture fermentations can improve chemical production from complex biosynthetic pathways over monocultures by distributing enzymes across multiple strains, thereby reducing metabolic burden, overcoming endogenous regulatory mechanisms, or exploiting natural traits of different microbial species. However, stabilizing and optimizing microbial sub-populations for maximal chemical production remains a major obstacle in the field. An optogenetic circuit, called OptoTA, is disclosed for regulating a toxin-antitoxin system, which enables tunability of, e.g., Escherichia coli growth using only blue light. With the disclosed system, one can control population ratios of co-cultures of, e.g., E. coli and Saccharomyces cerevisiae containing different metabolic modules of biosynthetic pathways. Results reveal that intermediate light duty cycles improve chemical production by establishing optimal co-culture populations.

The technology described herein is directed to regulated synthetic gene expression systems. In one aspect described herein are synthetic transcription factors (synTFs) comprising a DNA binding domain, a transcriptional effector domain, and a regulator protein. In other aspects described herein are gene expression systems comprising said synTFs and methods of treating diseases and disorders using said synTFs.

An auxin-inducible degron system kit that controls degradation of a target protein in a non-plant-derived eukaryotic cell, the kit containing a first nucleic acid that encodes a mutant TIR1 family protein having a mutation at an auxin-binding site, an auxin analog that has an affinity to the mutant TIR1 family protein and a second nucleic acid that encodes a degradation tag containing at least a part of an Aux/IAA family protein and having an affinity to a complex of the mutant TIR1 family protein and the auxin analog.

Described herein are a synthetic cascading bistable switches (Syn-CBS) circuit. In some aspects, the SynCBS circuit is single-strain circuit with two coupled self-activation modules to achieve two successive cell fate transitions. In other aspects, the SynCBS circuit is a two-strain circuit where the self-activation modules are divided in two cells instead of being expressed in a single cell. Also described are plasmids encoding the Syn-CBS circuits.

Provided are a method of overexpressing a target gene and/or a method of reprogramming cells, the method including steps of (a) introducing a vector into cells, into which vector a promoter and a target gene are inserted; and (b) applying an electromagnetic wave to the cells obtained in the step (a), and a method of treating a disease using the method.

When the method of overexpressing a target gene using the electromagnetic wave-reactive promoter of the present disclosure is used, it is possible to artificially regulate expression levels of desired target genes in a simple manner in vivo and in vitro and to regulate expression of the target genes until a desired predetermined time.

The present disclosure provides compositions and methods related to transcription factor systems. Such systems provide for modular and tunable protein expression driven by regulated transcriptional activity.

The present disclosure provides a system for the expression of target protein in conjunction with enhancer protein. The enhancer protein may be a viral protein that blocks nucleocytoplasmic transport. Also provided are polynucleotides, vectors, and cells comprising target protein and enhancer protein nucleic acid sequences.

The present disclosure relates to methods for transiently activating temperature-sensitive agents in one or more cells, for example by contacting one or more cells with a temperature-sensitive agent and transiently incubating the cells at a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. Additionally, the present disclosure relates to methods of contacting one or more cells in a subject with a temperature-sensitive agent and then lowering the subject's body temperature to a permissive temperature for inducing an activity of the temperature-sensitive agent in the cells. The disclosure also relates to methods of treating a subject with a temperature-sensitive therapeutic agent. In particular, the disclosure provides tools for temperature-sensitive delivery of ZSCAN4 nucleic acids and proteins to cells.