Disclosed is a technique for constructing optogenetic amplifier and inverter circuits utilizing transcriptional activator/repressor pairs, in which expression of the transcriptional activator or repressor, respectively, is controlled by light-controlled transcription factors. This system is demonstrated utilizing the quinic acid regulon system from Neurospora crassa, or Q System, a transcriptional activator/repressor system. This is also demonstrated utilizing the galactose regulon from Saccharomyces cerevisiae, or GAL System. Such optogenetic amplifier circuits enable multi-phase microbial fermentations, in which different light schedules are applied in each phase to dynamically control different metabolic pathways for the production of proteins, fuels or chemicals. The orthogonal nature of the Q and GAL systems enable the co-expression of amplifier and inverter circuits to simultaneously amplify and invert the response of light-controlled transcriptional controls over different sets of genes in the same cell.

The present disclosure provides expression constructs designed to provide for stable and/or inducible, tightly controlled production of genetically encoded payloads from engineered cells. These cassettes allow cells to be engineered to express genetically encoded payloads despite epigenetic silencing. As such, provided herein are expression systems for use in methods to engineer cells using CRISPR dCas9-activator systems such that expression of genetically encoded payloads (e.g., therapeutic proteins) can be optimized to overcome epigenetic silencing. In addition, provided herein are engineered cells comprising the expression systems.

Disclosed is an auto-induction regulatory system based on quorum sensing, comprising luxI, luxR and egfp, wherein, the promoter for controlling the expression of luxI and luxR is selected from P.sub.luxI, P.sub.BB or P.sub.J23100; the promoter for controlling the expression of egfp is selected from P.sub.luxI, P.sub.luxI(T-38C) or P.sub.luxI(C-77T). Also disclosed are an application of the auto-induction regulatory system based on quorum sensing in the automatic regulation of expression of a target gene of engineered Escherichia coli, as well as an application thereof in the preparation of alginate lyase and esterase. Further disclosed are a recombinant expression vector and a recombinant engineered bacterium comprising the auto-induction regulatory system based on quorum sensing.

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.

This invention relates to polynucleotides comprising a nucleotide sequence encoding a PPT1 polypeptide or a fragment thereof, vectors (viral or non-viral vectors) comprising the same, and methods of using the same for delivery of the open reading frame to a cell or a subject and to treat infantile neuronal lipofuscinosis (infantile Batten disease). The polynucleotides comprise an optimized CLN1 open reading frame.

The present disclosure relates to the use of an Epstein Barr virus origin of replication (oriP) or a functional fragment thereof in a protein expression construct to increase production of a protein of interest in mammalian cells. Also disclosed are protein expression constructs for increased production of antibodies in mammalian cells, and mammalian cells containing the expression constructs.

The present disclosure provides a nucleic acid sequence for regulating the transcription of a nucleic acid fragment of interest, wherein the nucleic acid sequence comprises at least 2 copies of TetO-operator sequences capable of binding to a transactivator rtTA regulatable by tetracycline or a derivative thereof, and 1 copy of minimal promoter sequence containing a TATA box sequence, and at least 1 copy of a CuO-operator sequence capable of binding to a transcription repressor CymR regulatable by cumate. The present disclosure also provides a vector and a host cell containing the nucleic acid sequence, and a method for inducing the expression of a nucleic acid fragment of interest in a host cell.

The disclosure provides methods and apparatus for producing 3-hydroxypropionic acid or a salt thereof, for removing 3-hydroxypropionic acid from aqueous solution (e.g., aqueous broth), and for using it to make various chemicals.

The present invention provides expression vectors for use in an inducible coexpression system, capable of controlled induction of expression of each gene product.

The present invention relates to a vaccine composition comprising an effective amount of a replication-competent controlled recombinant virus. Further encompassed are uses in immunization and methods of immunization employing compositions comprising a replication-competent controlled recombinant virus of the invention.