# US 2022/0056474 A1 · Host bacterium specific nanoparticle > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2022/0056474 A1 - Document ID: US20220056474A1 - Status: Published application - Kind: A1 - Application number: 17/275,401 - Filed: 2019-12-05 - Published: 2022-02-24 - Earliest claimed: 2018-12-27 - Assignees: National University Corporation Tokai National Higher Education and Research System - Applicants: National University Corporation Tokai National Higher Education and Research System - Inventors: Shoichi MITSUNAKA; Hiroki ANDO - Web page: https://patentgrep.com/patent/US20220056474A1 ## CPC - A61P 1/02: Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis - C12N 15/86: Viral vectors - A61K 35/74: Bacteria (therapeutic use of a bacterial protein A61K38/00) - A61K 35/76: Viruses; Subviral particles; Bacteriophages - A61P 31/04: Antibacterial agents - C12N 1/20: Bacteria; Culture media therefor - C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) - C12N 7/045: {Pseudoviral particles; Non infectious pseudovirions, e.g. genetically engineered} - C12N 2710/00032: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent - C12N 2795/10032: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) ## Abstract An object of the present invention is to provide a recombinant phage having high safety and excellent practicality and usefulness. Provided are a recombinant bacteriophage which is deprived of its proliferative capacity and can infect only once due to the fact that a bacteriophage genome in which a part of a virion constituent gene is deleted is stored in a head, and a method for preparing the same. In addition, provided are a recombinant bacteriophage which is deprived of its proliferative capacity and can infect only once due to the fact that a plasmid having a packaging site and encoding a target gene is stored in a head, and a method for preparing the same. ## Claims ### Claim 1 A method for preparing a host bacterium-specific nanoparticle, comprising the following steps (1) and (2): (1) a step of providing a recombinant vector ligated with a bacteriophage genome in which a part of a virion constituent gene is deleted; and (2) a step of causing a packaging reaction in the coexistence of the recombinant vector and a plasmid encoding the deleted virion constituent gene. ### Claim 2 (depends on 1) The preparation method according to claim 1, wherein the bacteriophage genome is provided as a plurality of fragments, and the recombinant vector is prepared by a seamless cloning method using the plurality of fragments and a linear vector. ### Claim 3 (depends on 2) The preparation method according to claim 2, wherein the seamless cloning method is gap-repair cloning or Gibson Assembly utilizing homologous recombination in yeast cells. ### Claim 4 (depends on 1) The preparation method according to claim 1, wherein the deleted virion constituent gene is a head gene or a tail gene. ### Claim 5 (depends on 1) The preparation method according to claim 1, wherein the deleted virion constituent gene is a tail gene. ### Claim 6 (depends on 1) The preparation method according to claim 1, wherein the bacteriophage genome is a genome of T7 phage. ### Claim 7 (depends on 1) The preparation method according to claim 1, wherein step (2) consists of the following steps (2-1) and (2-2): (2-1) a step of introducing the recombinant vector into a host bacterium carrying a plasmid that encodes the deleted virion constituent gene; and (2-2) a step of culturing the host bacterium after the introduction operation. ### Claim 8 (depends on 1) The method according to claim 1, further comprising the following step (3): (3) a step of recovering a bacteriophage generated by the packaging reaction. ### Claim 9 A host bacterium-specific nanoparticle, consisting of a recombinant bacteriophage that comprises a head and a tail, and has an ability to infect but no ability to re-infect a host bacterium, the head having stored therein a bacteriophage genome in which a part of a virion constituent gene is deleted. ### Claim 10 (depends on 9) The host bacterium-specific nanoparticle according to claim 9, wherein the deleted virion constituent gene is a head gene or a tail gene. ### Claim 11 (depends on 9) The host bacterium-specific nanoparticle according to claim 9, wherein the deleted virion constituent gene is a tail gene. ### Claim 12 (depends on 9) The host bacterium-specific nanoparticle according to claim 9, wherein the bacteriophage genome is a genome of T7 phage. ### Claim 13 (depends on 9) An antibacterial agent comprising the host bacterium-specific nanoparticle according to claim 9 as an active ingredient. ### Claim 14 (depends on 13) A composition comprising the antibacterial agent according to claim 13. ### Claim 15 (depends on 14) The composition according to claim 14, which is a pharmaceutical, disinfectant, cleaning agent, or oral composition against bacterial infections. ### Claim 16 A method for preparing a host bacterium-specific nanoparticle, comprising the following steps (1) and (2): (1) a step of providing a recombinant vector ligated with a bacteriophage genome in which a packaging site is deleted; and (2) a step of causing a packaging reaction in the coexistence of the recombinant vector and a plasmid having the deleted packaging site and encoding a target gene. ### Claim 17 (depends on 16) The preparation method according to claim 16, wherein the bacteriophage genome is provided as a plurality of fragments, and the recombinant vector is prepared by a seamless cloning method using the plurality of fragments and a linear vector. ### Claim 18 (depends on 17) The preparation method according to claim 17, wherein the seamless cloning method is gap-repair cloning utilizing homologous recombination in yeast cells. ### Claim 19 (depends on 16) The preparation method according to claim 16, wherein the bacteriophage genome is a genome of T7 phage. ### Claim 20 (depends on 16) The preparation method according to claim 16, wherein the target gene is one or more genes selected from the group consisting of a marker gene, a reporter gene, an enzyme gene, a gene for genome editing, a gene encoding an antibacterial peptide, an antibacterial gene, and a group of genes constituting a synthetic gene circuit. ### Claim 21 (depends on 16) The method according to claim 16, wherein step (2) consists of the following steps (2-1) and (2-2): (2-1) a step of introducing the recombinant vector into a host bacterium carrying a plasmid that has the deleted packaging site and encodes the target gene; and (2-2) a step of culturing the host bacterium after the introduction operation and then lysing the host bacterium. ### Claim 22 (depends on 16) The method according to claim 16, further comprising the following step (3): (3) a step of recovering a bacteriophage generated by the packaging reaction. ### Claim 23 A host bacterium-specific nanoparticle, consisting of a recombinant bacteriophage that comprises a head and a tail, and has an ability to infect but no ability to re-infect a host bacterium, the head having stored therein a plasmid having a packaging site and encoding the target gene. ### Claim 24 (depends on 23) The host bacterium-specific nanoparticle according to claim 23, wherein the bacteriophage genome is a genome of T7 phage. ### Claim 25 (depends on 23) The host bacterium-specific nanoparticle according to claim 23, wherein the target gene is one or more genes selected from the group consisting of a marker gene, a reporter gene, an enzyme gene, a gene for genome editing, a gene encoding an antibacterial peptide, an antibacterial gene, and a group of genes constituting a synthetic gene circuit. ### Claim 26 (depends on 23) A composition for transduction, comprising the host bacterium-specific nanoparticle according to claim 23 as an active ingredient. ### Claim 27 A recombinant bacteriophage comprising a head and a tail, and having an ability to infect but no ability to re-infect a host bacterium, the head having stored therein a bacteriophage genome in which a part of a virion constituent gene is deleted. ### Claim 28 (depends on 27) The recombinant bacteriophage according to claim 27, wherein the recombinant bacteriophage is a virulent phage. ### Claim 29 (depends on 27) A pharmaceutical composition comprising the recombinant bacteriophage according to claim 27 and an excipient. ### Claim 30 (depends on 29) The pharmaceutical composition according to claim 29 for the prevention or treatment of bacterial infections.