# US 11,078,490 B2 · Optimized vector for delivery in microbial populations > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,078,490 B2 - Document ID: US11078490B2 - Status: Granted - Kind: B2 - Application number: 17/017,111 - Filed: 2020-09-10 - Published: 2020-12-24 - Granted: 2021-08-03 - Earliest claimed: 2017-02-03 - Assignees: Eligo Bioscience; Institut Pasteur - Applicants: Eligo Bioscience; Institut Pasteur - Inventors: Antoine Decrulle; Jesus Fernandez Rodriguez; Xavier Duportet; David Bikard - Web page: https://patentgrep.com/patent/US11078490B2 ## CPC - C12N 15/73: Expression systems using phage (lambda) regulatory sequences - A61K 35/76: Viruses; Subviral particles; Bacteriophages - A61P 31/04: Antibacterial agents - 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 15/70: Vectors or expression systems specially adapted for E. coli - C12N 15/74: Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora - C12N 2795/10332: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent - C12N 2800/10: Plasmid DNA - C12N 2800/101: for bacteria - Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract The present invention relates to a vector, preferably included in a delivery vehicle, comprising no more than 100, preferably no more than 10, restriction sites recognized by the restriction enzymes encoded by each bacterium of a group of bacteria of interest. The invention also relates to the use of said vector, preferably included in a delivery vehicle, as a drug, especially in the treatment of a disease in a patient in need thereof. ## Claims ### Claim 1 An engineered bacteriophage or packaged phagemid, for DNA delivery into Klebsiella pneumoniae strains, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove one or more of the restriction sites selected from the group consisting of CAGNNNNNNCGT (SEQ ID NO: 93); GAAYNNNNNNNCTGG (SEQ ID NO: 94); CGANNNNNNNNTGCC (SEQ ID NO: 95); ACGNNNNNGTTG (SEQ ID NO: 96) and CGCATC corresponding to restriction enzymes encoded by Klebsiella pneumoniae strains. ### Claim 2 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites CAGNNNNNNCGT (SEQ ID NO: 93); GAAYNNNNNNNCTGG (SEQ ID NO: 94); CGANNNNNNNNTGCC (SEQ ID NO: 95); ACGNNNNNGTTG (SEQ ID NO: 96) and CGCATC corresponding to restriction enzymes encoded by Klebsiella pneumoniae strains. ### Claim 3 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction site CAGNNNNNNCGT (SEQ ID NO: 93). ### Claim 4 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction site GAAYNNNNNNNCTGG (SEQ ID NO: 94). ### Claim 5 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction site CGANNNNNNNNTGCC (SEQ ID NO: 95). ### Claim 6 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction site ACGNNNNNGTTG (SEQ ID NO: 96). ### Claim 7 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction site CGCATC. ### Claim 8 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites CAGNNNNNNCGT (SEQ ID NO: 93) and CGCATC. ### Claim 9 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites CAGNNNNNNCGT (SEQ ID NO: 93), GAAYNNNNNNNCTGG (SEQ ID NO: 94) and CGCATC. ### Claim 10 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites CAGNNNNNNCGT (SEQ ID NO: 93), GAAYNNNNNNNCTGG (SEQ ID NO: 94), CGANNNNNNNNTGCC (SEQ ID NO: 95) and CGCATC. ### Claim 11 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites GAAYNNNNNNNCTGG (SEQ ID NO: 94) and CGCATC. ### Claim 12 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites CGANNNNNNNNTGCC (SEQ ID NO: 95) and CGCATC. ### Claim 13 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites CAGNNNNNNCGT (SEQ ID NO: 93) and GAAYNNNNNNNCTGG (SEQ ID NO: 94). ### Claim 14 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites CAGNNNNNNCGT (SEQ ID NO: 93) and CGANNNNNNNNTGCC (SEQ ID NO: 95). ### Claim 15 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites GAAYNNNNNNNCTGG (SEQ ID NO: 94) and CGANNNNNNNNTGCC (SEQ ID NO: 95). ### Claim 16 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein said engineered bacteriophage or packaged phagemid has been genetically engineered to remove the restriction sites GAAYNNNNNNNCTGG (SEQ ID NO: 94), CGANNNNNNNNTGCC (SEQ ID NO: 95) and CGCATC. ### Claim 17 (depends on 1) The engineered bacteriophage or packaged phagemid according to claim 1, wherein the bacteriophage or packaged phagemid for DNA delivery into Klebsiella pneumoniae strain is derived from a bacteriophage selected from the group consisting of AIO-2, KI4B, K16B, K19, (syn=K19), K114, K115, K121, K128, K129, KI32, K133, K135, K1106B, K1171B, K1181B, K1832B, AIO-I, AO-I, AO-2, AO-3, FC3-10, K, K11, (syn=KI1), K12, (syn=K12), K13, (syn=K13), (syn=Kl 70/11), K14, (syn=K14), K15, (syn=K15), K16, (syn=K16), K17, (syn=K17), K18, (syn=K18), K119, (syn=K19), K127, (syn=K127), K131, (syn=K131), K135, K1171B, II, VI, IX, CI-I, K14B, K18, K111, K112, K113, K116, K117, K118, K120, K122, K123, K124, K126, K130, K134, K1106B, KIi65B, K1328B, KLXI, K328, P5046, 11, 380, III, IV, VII, VIII, FC3-11, K12B, (syn=K12B), K125, (syn=K125), K142B, (syn=K142), (syn=K142B), K1181B, (syn=KI1 81), (syn=K1181B), K1765/!, (syn=K1765/1), K1842B, (syn=K1832B), K1937B, (syn=K1937B), Ll, φ28, 7, 231, 483, 490, 632, 864/100, KP01K2, Kl l, Kpn5, KP34, and JDOO1. ### Claim 18 (depends on 1) A pharmaceutical or veterinary composition, comprising an engineered bacteriophage or a packaged phagemid according to claim 1. ### Claim 19 (depends on 18) A method of treating a subject infected with pathogenic or virulent bacteria comprising the administration of the pharmaceutical or veterinary composition of claim 18. ### Claim 20 (depends on 19) The method of claim 19, wherein the subject is infected with Klebsiella pneumoniae.