# US 2024/0409588 A1 · Methods and products for genetic engineering > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2024/0409588 A1 - Document ID: US20240409588A1 - Status: Published application - Kind: A1 - Application number: 18/742,986 - Filed: 2024-06-13 - Published: 2024-12-12 - Earliest claimed: 2015-10-20 - Assignees: Institut National De La Sante Et De La Recherche Medicale (Inserm); Centre National de la Recherche Scientifique (CNRS); Ecole Normale Superieure De Lyon; Universite Claude Bernard Lyon 1 - Applicants: Institut National De La Sante Et De La Recherche Medicale (INSERM); Centre National De La Recherche Scientifique (CNRS); Ecole Normale Superieure De Lyon; Universite Claude Bernard Lyon 1 - Inventors: Théophile OHLMANN; Philippe MANGEOT; Emiliano RICCI - Web page: https://patentgrep.com/patent/US20240409588A1 ## CPC - C07K 14/005: from viruses - 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 9/22: Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} - C12N 9/224: {Class 1 CAS enzyme complex, e.g. multi-enzyme} - C12N 15/102: {Mutagenizing nucleic acids} - C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} - C12N 15/90: Stable introduction of foreign DNA into chromosome - C07K 2319/00: Fusion polypeptide - C07K 2319/43: containing a FLAG-tag - C07K 2319/50: containing protease site - C12N 2310/20: involving clustered regularly interspaced short palindromic repeats [CRISPR] - C12N 2740/10023: Virus like particles [VLP] - C12N 2740/10042: virus or viral particle as vehicle, e.g. encapsulating small organic molecule - C12N 2740/13023: Virus like particles [VLP] - C12N 2740/13042: virus or viral particle as vehicle, e.g. encapsulating small organic molecule - C12N 2800/80: Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract The present invention relates to a virus-derived particle comprising one or more Cas protein(s), as well as to kits and methods using the same for altering a target nucleic acid. ## Claims ### Claim 1 A retrovirus-derived particle comprising one or more free Cas protein(s), wherein the one or more free Cas protein(s) is inside of the particle, wherein the one or more free Cas protein(s) is the result of cleavage of a cleavable fusion protein comprising a proteolysis cleavage site located between one or more Cas proteins and a retroviral gag protein, and wherein the one or more free Cas protein(s) is complexed with one or more CRISPR-Cas guide RNA(s). ### Claim 2 (depends on 1) The retrovirus-derived particle of claim 1, wherein the free Cas protein is Cas9 or an homolog or a derivative thereof. ### Claim 3 (depends on 1) The retrovirus-derived particle of claim 1, wherein the free Cas protein is Cas9. ### Claim 4 (depends on 3) The retrovirus-derived particle of claim 3, wherein the free Cas9 is from Streptococcus pyogenes (spCas9). ### Claim 5 (depends on 1) The retrovirus-derived particle of claim 1, which is a lentivirus-derived particle. ### Claim 6 (depends on 3) The retrovirus-derived particle of claim 3, which is a lentivirus-derived particle. ### Claim 7 (depends on 4) The retrovirus-derived particle of claim 4, which is a lentivirus-derived particle. ### Claim 8 (depends on 1) The retrovirus-derived particle of claim 1, wherein the retrovirus-derived particle is substantially devoid of any encoding nucleic acid. ### Claim 9 (depends on 1) The retrovirus-derived particle of claim 1, wherein the retrovirus-derived particle is devoid of any nucleic acid encoding a viral protein. ### Claim 10 (depends on 1) The retrovirus-derived particle of claim 1, wherein the CRISPR-Cas system guide RNAs comprise: (a) a first CRISPR-Cas system guide RNA that hybridizes with a first target sequence of a target nucleic acid, and (b) a second CRISPR-Cas system guide RNA that hybridizes with a second target sequence of said target nucleic acid. ### Claim 11 (depends on 1) The retrovirus-derived particle of claim 1, wherein the retrovirus-derived particle is selected from the group consisting of Moloney murine leukemia virus-derived vector particles, Bovine immunodeficiency virus-derived particles, Simian immunodeficiency virus-derived vector particles, Feline immunodeficiency virus-derived vector particles, Human immunodeficiency virus-derived vector particles, Equine infection anemia virus-derived vector particles, Caprine arthritis encephalitis virus-derived vector particle, and Baboon endogenous virus-derived vector particles. ### Claim 12 (depends on 4) The retrovirus-derived particle of claim 4, wherein the retrovirus-derived particle is selected from the group consisting of Moloney murine leukemia virus-derived vector particles, Bovine immunodeficiency virus-derived particles, Simian immunodeficiency virus-derived vector particles, Feline immunodeficiency virus-derived vector particles, Human immunodeficiency virus-derived vector particles, Equine infection anemia virus-derived vector particles, Caprine arthritis encephalitis virus-derived vector particle, and Baboon endogenous virus-derived vector particles. ### Claim 13 (depends on 12) The retrovirus-derived particle of claim 12, wherein the retrovirus-derived particle is a Moloney murine leukemia virus-derived vector particle. ### Claim 14 (depends on 12) The retrovirus-derived particle of claim 12, wherein the retrovirus-derived particle is a Human immunodeficiency virus-derived vector particle. ### Claim 15 (depends on 12) The retrovirus-derived particle of claim 12, where the retrovirus-derived particle is a Baboon endogenous virus-derived vector particle. ### Claim 16 (depends on 1) The retrovirus-derived particle of claim 1, wherein the retrovirus-derived particle further comprises a viral protease. ### Claim 17 (depends on 1) The retrovirus-derived particle of claim 1, wherein the retrovirus-derived particle is devoid of any viral protease. ### Claim 18 (depends on 1) The retrovirus-derived particle of claim 1, wherein the retroviral gag protein is derived from Moloney murine leukemia virus. ### Claim 19 (depends on 1) The retrovirus-derived particle of claim 1, wherein the retroviral gag protein is derived from Human immunodeficiency virus. ### Claim 20 (depends on 1) The retrovirus-derived particle of claim 1, wherein the retroviral gag protein is derived from Baboon endogenous virus.