In some aspects, the disclosure relates to recombinant adeno-associated viruses (rAAVs) comprising a nucleic acid encoding a fusion protein comprising a DNA-binding domain and a transcriptional regulator domain and methods of using the same. In some embodiments, expression of the fusion protein results in modified expression of a target gene in a cell.

Described herein are transcriptional relay systems useful for reducing background signal in protein expression and reporter assays. These systems utilize a nucleic acid system wherein a promoter sequence controls expression of a synthetic transcription factor that activates transcription of a reporter molecule.

Disclosed herein include circuits, compositions, nucleic acids, populations, systems, and methods enabling single circuits to generate multiple molecularly and functionally distinct states that are each stable across multiple cell division cycles. Synthetic circuits provided herein can stably exist in multiple distinct states characterized by differences in the concentrations and expression levels of its components. In the absence of changes to the external environment, each of these states can be stable. In some embodiments, transcription factors provided herein activate when dimerized, and show much weaker activity as monomers. In some embodiments, each transcription factor homodimer activates expression of its own gene. In some embodiments, transcription factors can form mixed heterodimers with one another that do not strongly activate any genes in the circuit. Different embodiments of the synthetic circuits provided herein can use different numbers of transcription factors to produce a growing number of stable states.

This present patent application relates to a method for controlling a targeted gene expression with an environmental trigger useful for manufacturing a natural product or an analogue thereof biologically. In particular, the present invention discloses an expression system comprises a gene expressing for a fusion protein of elastin-like polypeptides (ELPs) and a transcription factor, wherein a targeted gene expression is regulated by the phase change of the fusion protein initiated by an environmental trigger, including changes of temperature, pH value, ionic strength or a combination thereof. The method, the expression system and their products are within the scope of this invention.

Engineered CRISPR-Cas9 nucleases with altered and improved PAM specificities and their use in genomic engineering, epigenomic engineering, and genome targeting.

The invention relates to nucleic acid molecules, vectors and plasmids comprising AAV cap genes and rep genes, wherein the cap and rep genes are both operably-associated with an inducible promoter which comprises one or more cumate operator (CuO) sequences. The invention further relates to producer and packaging cell lines which are useful in the production of Adeno-Associated Virus (AAV) particles.

This disclosure concerns compositions and methods for increasing the expression of a polynucleotide of interest. Some embodiments concern novel transactivation polypeptides and variants thereof that have been identified in plants, and methods of using the same. Particular embodiments concern the use of at least one DNA-binding polypeptide in a fusion protein to target at least one transactivation polypeptide or variant thereof to a specific binding site on a nucleic acid comprising the polynucleotide of interest, such that its expression may be increased.

Provided herein are nucleic acid molecules, vectors, and recombinant AAV comprising an inducible gene expression system. The system includes a transgene encoding a gene product operably linked to expression control sequences comprising a promoter; an activation domain comprising a canine or feline transactivation domain and a FKBP12-rapamycin binding (FRB) domain of canine or feline FKBP12-rapamycin-associated protein (FRAP); a DNA binding domain comprising a zinc finger homeodomain (ZFHD) and one, two or three FK506 binding protein domain (FKBP) subunit genes; and at least 8 copies of the binding site for ZFHD (8XZFHD) followed by a minimal IL2 promoter. The presence of an effective amount of a rapamycin or a rapalog induces expression of the transgene in a host cell.

Engineered CRISPR-Cas9 nucleases with altered and improved PAM specificities and their use in genomic engineering, epigenomic engineering, and genome targeting.

Described herein are novel systems for targeting, editing or manipulating DNA in a cell, using novel M-SmallCas9 nucleases and variants thereof. The M-SmallCas9 nucleases are derived from wildtype or parental small type II CRISPR Cas9 endonucleases, and display improved fidelity compared to parental type II CRISPR Cas9 enzymes in combination with a simple PAM sequences and are small endonuclease size.