Provided is a modified bacteriophage capable of infecting a target bacterium, which bacteriophage includes an α/β small acid-soluble spore protein (SASP) gene encoding a SASP which is toxic to the target bacterium, wherein the SASP gene is under the control of a constitutive promoter which is foreign to the bacteriophage and the SASP gene.

The present disclosure relates generally to methods and compositions for transferring a genetic circuit from one prokaryotic cell (“donor cell”) to another prokaryotic cell (“recipient cell” or “target cell” which are used interchangeably herein). More specifically, the present disclosure relates to prokaryotic donor cells comprising (i) a genetic circuit of interest and (ii) one or more expressed transcriptional repressor proteins and the use of said donor cells in the efficient transfer of the genetic circuit into a prokaryotic recipient cell. The genetic circuit includes nucleic acid sequences encoding a RNA molecule or protein of interest.

The present invention concerns a production bacterial cell for producing phage particles or phage-derived delivery vehicles, said production bacterial cell stably comprising at least one phage structural gene(s) and at least one phage DNA packaging gene(s), said phage structural gene(s) and phage DNA packaging gene(s) being derived from a first type of bacteriophage, wherein the expression of at least one of said phage structural gene(s) and/or at least one of said phage DNA packaging gene(s) in said production bacterial cell is controlled by at least one induction mechanism, and wherein said production bacterial cell is from a bacterial species or strain different from the bacterial species or strain from which said first type of bacteriophage comes and/or that said first type of bacteriophage targets.

The present invention is related to the field of CRISPR-Cas9 gene editing platforms. In particular, the present invention has identified Type II-C Cas9 anti-CRISPR (Acr) inhibitors that control Cas9 gene editing activity. Co-administration of such Acr inhibitors may provide an advantageous adjunct in permitting safe and practical biological therapeutics through spatial or temporal control of Cas9 activity; controlling Cas9-based gene drives in wild populations to reduce the ecological consequences of such forced inheritance schemes; and contributing to general research into various biotechnological, agricultural, and medical applications of gene editing technologies.

The methods described herein are for treating infections in individuals having cancer and who are receiving cancer immunotherapy, preferably employing a CRISPR system to selectively kill or reduce the numbers of pathogenic bacteria within the individual and thereafter, administering an immune checkpoint inhibitor thereto. In particular embodiments, the pathogenic bacteria is one of E. coli, Pseudomonas aeruginosa, Klebsiella bacteria, Staphylococcus aureus; Streptoccocus; Salmonella; Shigella; Mycobacterium tuberculosis; Enterococcus; Clostridium; Neisseria gonnorrhoea; Acinetobacter baumannii; and Campylobacter bacteria and the checkpoint inhibitor is selected from the group consisting of nivolumab, pembrolizumab, pidilizumab, AMP-224, AMP-514, STI-A1110, TSR-042, RG-7446, BMS-936559, MEDI-4736, MSB-0020718C, AUR-012 and STI-A1010. Further embodiments include enhancing the growth of a second bacteria in the individual, such bacteria including Akkermansia, Bacteroides, Bifidobacterium, Enterococcus, Fusobacterium, Coprococcus, LactoBacillus, Propionibacterium, Ruminococcus, Veillonella, Prevotella, and F. prausnitzii. The CRISPR system may include Cas9, Cpf1 and Cas3, and may be delivered using a bacteriophage.

The present invention concerns a production bacterial cell for producing lytic phage particles or lytic phage-derived delivery vehicles, said production bacterial cell stably comprising at least one phage structural genes and at least one phage DNA packaging genes, said phage structural gene(s) and phage DNA packaging gene(s) being derived from a lytic bacteriophage, wherein the expression of at least one of said phage structural genes and/or at least one of said phage DNA packaging gene(s) in said production bacterial cell is controlled by an induction mechanism.

The present disclosure relates generally to methods and compositions for transferring a genetic circuit from one prokaryotic cell (“donor cell”) to another prokaryotic cell (“recipient cell” or “target cell” which are used interchangeably herein). More specifically, the present disclosure relates to prokaryotic donor cells comprising (i) a genetic circuit of interest and (ii) one or more expressed transcriptional repressor proteins and the use of said donor cells in the efficient transfer of the genetic circuit into a prokaryotic recipient cell. The genetic circuit includes nucleic acid sequences encoding a RNA molecule or protein of interest.

Modified bacteriophage, uses thereof, and compositions containing the modified bacteriophage are described. The compositions are useful for human treatment and may treat various conditions, including bacterial infections.

Provided herein are methods and compositions for killing a target bacterium. Also disclosed are engineered bacteriophages.

A method of inhibiting growth of a bacterium including steps of: a) delivering a polynucleotide including a sequence of SEQ ID NO: 1 or a homologue thereof to the bacterium, and b) contacting the bacterium with an effective amount of an antibiotic; wherein the bacterium is Pseudomonas aeruginosa. A method of treating a subject suffering from an infection caused by a bacterium which is Pseudomonas aeruginosa and the method including steps of: i) delivering said polynucleotide to a tissue infected by the bacterium or a bacterial cell in the subject; and ii) administering an effective amount of an antibiotic to the subject. A composition and a recombinant plasmid containing said polynucleotide.