A nucleic acid molecule is uniquely designed and encodes an entire CRISPRi or CRISPRa system, while being sized for packaging within a single adeno-associated virus (AAV) vector. Examples of the nucleic acid molecule include about 4600 to 4700 base pairs. Examples of the nucleic acid molecule can include a nucleotide encoding a Cas polypeptide; a nucleotide encoding a repressor or an activator domain attached to the nucleotide encoding the Cas polypeptide via a linker; a first promoter operably connected to the nucleotide encoding the repressor or activator domain or the nucleotide encoding the Cas polypeptide; a nucleotide encoding an alpha-helical connecting the nucleotide encoding the Cas polypeptide to a nuclear localization signal (NLS); and a second promoter operably connected to a guide RNA (gRNA).

The invention relates to the production and use of Cas-encoding sequences and vectors comprising these. Aspects of the invention provide products, vectors, delivery vehicles, uses and methods for producing Cas-encoding sequences in bacterial or archaeal cells.

The invention relates to the production and use of Cas-encoding sequences and vectors comprising these. Aspects of the invention provide products, vectors, delivery vehicles, uses and methods for producing Cas-encoding sequences in bacterial or archaeal cells.

The present disclosure discloses a Bacillus subtilis efficiently-induced expression system based on an artificial series promoter, and belongs to the technical field of gene engineering. According to the present disclosure, an efficient artificial series constitutive promoter is used, and by compositely designing the promoter and elements (a repressor and a binding site thereof) relevant to an operon, the activity of the constitutive promoter is regulated and controlled by an inducer to finally construct the B. subtilis efficiently-induced expression system induced by the inducer. The result indicates that compared with a P.sub.43 strongly constitutive promoter, the activity of the artificial series promoter in the system is about 15 times higher. The activity of the promoter can be accurately controlled by adding different concentrations of inducers. Therefore, the efficient expression system is simple in structure, high in activity and strict in regulating and controlling and has wide application prospects in heterologous protein efficient expression and synthetic biology research.

Disclosed herein are optogenetic circuits for the bacterium Escherichia coli that induce gene expression in darkness and repress it under blue light. Applying them to metabolic engineering improves chemical production compared to chemically induced controls in light-controlled fermentations. More particularly, these circuits can be used to control protein production with light. The system and method use light as a suitable alternative to chemical induction for microbial production of chemicals and proteins.

The present invention provides a cell which expresses a chimeric antigen receptor (CAR) or an engineered T-cell receptor (TCR), the cell comprising a nucleotide sequence of interest (NOI) which is selectively expressed by the cell depending on: i) the differentiation/exhaustion state of the cell; or ii) the presence of an environmental metabolite in the microenvironment of the cell.

The present disclosure provides a system for regulating expression of a target polynucleotide in a cell. The system may comprise a chimeric polypeptide comprising a gene modulating polypeptide fused in-frame with a heterologous nuclear localization domain. The heterologous nuclear localization domain may be operable to translocate the chimeric polypeptide to a cell nucleus upon activation by an active cellular signaling pathway. The cellular signaling pathway may be inducible in response to an extracellular signal. In response to the extracellular signal, the chimeric polypeptide may localize to the cell nucleus and the gene modulating polypeptide may regulate expression of a target polynucleotide in the cell nucleus.

The invention relates to the production and use of Cas-encoding sequences and vectors comprising these. Aspects of the invention provide products, vectors, delivery vehicles, uses and methods for producing Cas-encoding sequences in bacterial or archaeal cells.

The present disclosure relates to regulatable expression vectors for systematically controlling and silencing expression of a therapeutic molecule and uses thereof, e.g. treating ocular disorders such as choroidal neovascularisation.

Expression vectors ideal for use in vaccinating individuals against disease based on vaccinia virus and other chordopoxviruses having high expression of recombinant genes and low expression of vector genes in target animals, and low expression of recombinant genes and high expression of vector genes in cells used for propagation.