The present disclosure provides compositions and methods for introducing or enhancing Aca activity in prokaryotic cells. The provided compositions and methods can be used to inhibit Acr activity in prokaryotic cells, thereby enhancing endogenous or exogenous CRISPR-Cas activity. Cells, polynucleotides, plasmids, phage, and other elements for practicing the present methods are also provided.

Provided herein are, inter alia, compositions, methods, and systems for selectively modulating an activity of CRISPR-based systems, including CRISPR-based genome editing systems and CRISPR-based gene regulation in a target cell. Also provided are compositions, methods, and systems for controlling a gene regulation circuit or building dynamic pulsatile gene regulation circuits in a target cell, e.g., a mammalian cell by using CRISPR-based genome editing tools.

Provided herein are encapsulated cells that comprise a microbial cell modified to express a cytolytic enzyme and encapsulated with one or more layers of a cationic polymer and one or more layers of an anionic polymer that may be used in cell-free protein synthesis. To this end, methods of utilizing such encapsulated cells in synthesizing a target protein, degrading a contaminant in a contaminated environment and detecting an analyte in a sample are also disclosed herein. Further provided herein, are methods of producing such encapsulated cells.

The present disclosure relates generally to bacterial delivery vehicles for use in efficient transfer of a desired payload into a target bacterial cell. More specifically, the present disclosure relates to bacterial delivery vehicles with desired host ranges based on the presence of a chimeric receptor binding protein (RBP) composed of a fusion between the N-terminal region of a RBP derived from a lambda-like bacteriophage and the C-terminal region of a different RBP, and/or the presence of an engineered branched receptor binding multi-subunit polypeptides (“branched-RBP”).

Some aspects of this disclosure provide methods for phage-assisted continuous evolution (PACE) of proteases. Some aspects of this invention provide methods for evaluating and selecting protease inhibitors based on the likelihood of the emergence of resistant proteases as determined by the protease PACE methods provided herein. Some aspects of this disclosure provide strategies, methods, and reagents for protease PACE, including fusion proteins for translating a desired protease activity into a selective advantage for phage particles encoding a protease exhibiting such an activity and improved mutagenesis-promoting expression constructs. Evolved proteases that recognize target cleavage sites which differ from their canonical cleavage site are also provided herein.

Bacteriophage against Geobacillus are provided, and methods of making and using the bacteriophage also are provided.

The present invention relates to methods of treating or preventing a bacterial disease or infection, antibacterial compositions, and antibacterial surfaces, including an isolated polypeptide comprising an enzymatically active domain (EAD) of a Bacillus bacteriophage endolysin.

The present invention provides for a nucleic acid encoding a bacteriophage genome comprising a unique n-mer barcode inserted in a non-essential location or gene location within the bacteriophage genome, or a bacteriophage comprising the nucleic acid thereof

Bacteriophage are provided, and methods of making and using the bacteriophage also are provided.

The present invention relates to a polynucleotide encoding a fusion polypeptide comprising an anti-CRISPR (Acr) polypeptide, wherein said fusion polypeptide further comprises a receptor domain changing conformation upon reception of a stimulus. The present invention also relates to a vector comprising the polynucleotide of the present invention, to a bipartite Acr polypeptide comprising a first partial Acr polypeptide comprising amino acids corresponding to amino acids 10 to 62 of SEQ ID NO: 1, and a second partial Acr polypeptide comprising amino acids corresponding to amino acids 67 to 77 of SEQ ID NO: 1, and to a host cell comprising the aforesaid polynucleotide compounds. The present invention also relates to the said compounds for use in medicine, in particular for use in treatment and/or prevention of genetic disease, neurodegenerative disease, cancer, and/or infectious disease. Moreover, the present invention also relates to a kit, methods, and uses related thereto.