# US 2022/0033781 A1 · Artificial recombinant chromosome and use thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2022/0033781 A1 - Document ID: US20220033781A1 - Status: Published application - Kind: A1 - Application number: 16/626,816 - Filed: 2019-11-12 - Published: 2022-02-03 - Earliest claimed: 2019-04-12 - Assignees: Humab LLC - Applicants: Humab Co., Ltd. - Inventors: Chang Kyu Oh; Soon-Ik Park; Ho Jin Kang; Ae Jin Choi - Web page: https://patentgrep.com/patent/US20220033781A1 ## CPC - C12N 5/166: {resulting from interspecies fusion} - A01K 67/0278: Knock-in vertebrates, e.g. humanised vertebrates - C12N 5/0696: {Artificially induced pluripotent stem cells, e.g. iPS} - C12N 15/8509: {for producing genetically modified animals, e.g. transgenic} - A01K 2207/15: Humanized animals - A01K 2217/072: maintaining or altering function, i.e. knock in - A01K 2227/105: Murine - A01K 2267/01: Animal expressing industrially exogenous proteins - C12N 2800/30: Vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS ## Abstract An artificial recombinant chromosome produced by recombination of two or more chromosomes and the production of a transgenic animal using a cell including the same are disclosed. A method for producing a cell including one or more artificial recombinant chromosomes includes preparing a first targeted cell and a second targeted cell, producing one or more microcells using the second targeted cell, producing a fusion cell using the first targeted cell and the one or more microcells, and producing a cell including an artificial recombinant chromosome by treating the fusion cell with site specific recombinase (SSR). ## Claims ### Claim at position 1 1-12. (canceled) ### Claim 13 A method for producing a cell including one or more artificial recombinant chromosomes, the method comprising i) preparing a first targeted cell and a second targeted cell, wherein the first targeted cell comprises a first targeted chromosome, and the second targeted cell comprises a second targeted chromosome, wherein the first targeted chromosome includes a first part, a first recombinase recognition sequence (a first RRS), a first fragment, a second recombinase recognition sequence (a second RRS) and a second part, herein, the first part includes one of telomere ends located on both sides of the first targeted chromosome, and the second part includes the other one of telomere ends located on both sides of the first targeted chromosome, herein, the first fragment is located between the first RRS and the second RRS, wherein the second targeted chromosome includes a third part, a third recombinase recognition sequence (a third RRS), a second fragment, a fourth recombinase recognition sequence (a fourth RRS) and a fourth part, herein, the third part includes one of telomere ends located on both sides of the second targeted chromosome, and the fourth part includes the other one of telomere ends located on both sides of the second targeted chromosome, herein, the second fragment is located between the third RRS and the fourth RRS; ii) producing one or more microcells using the second targeted cell, wherein the one or more microcells comprise the second targeted chromosome or a fragment thereof, herein, the fragment of the second targeted chromosome includes the third RRS, the second fragment and the fourth RRS; iii) producing a fusion cell using the first targeted cell and the one or more microcells, wherein the fusion cell comprises the first targeted chromosome and the second targeted chromosome, or the first targeted chromosome and the fragment of the second targeted chromosome; and iv) producing a cell including an artificial recombinant chromosome by treating the fusion cell with site specific recombinase (SSR), wherein the SSR induces a recombination by recognizing a pairing of the first RRS present in the first targeted chromosome and the third RRS present in the second targeted chromosome, and a pairing of the second RRS present in the first targeted chromosome and the fourth RRS present in the second targeted chromosome, herein, the first fragment present in the first targeted chromosome is exchanged for the second fragment present in the second targeted chromosome by the recombination, thereby, a first artificial recombinant chromosome including the first part, the second fragment and the second part is produced. ### Claim 14 (depends on 13) The method of claim 13, wherein the cell including one or more artificial recombinant chromosomes further comprises a second artificial recombinant chromosome, herein, the second artificial recombinant chromosome includes the third part, the first fragment and the fourth part. ### Claim 15 (depends on 13) The method of claim 13, wherein the first part includes a centromere of the first targeted chromosome. ### Claim 16 (depends on 13) The method of claim 13, wherein the third part includes a centromere of the second targeted chromosome. ### Claim 17 (depends on 13) The method of claim 13, wherein the first fragment includes a centromere of the first targeted chromosome, wherein the second fragment includes a centromere of the second targeted chromosome. ### Claim 18 (depends on 13) The method of claim 13, wherein the first RRS is one selected from a loxP and a loxP variant, wherein the third RRS is one selected from a loxP and a loxP variant, herein, the first RRS is capable of pairing with the third RRS. ### Claim 19 (depends on 13) The method of claim 13, wherein the second RRS is one selected from a loxP and a loxP variant, wherein the fourth RRS is one selected from a loxP and a loxP variant, herein, the second RRS is capable of pairing with the fourth RRS. ### Claim 20 (depends on 18) The method of claim 18, wherein the SSR is a Cre recombinase. ### Claim 21 (depends on 13) The method of claim 13, wherein the first RRS is one selected from FRT, attP, attB, ITR and variants thereof, wherein the third RRS is one selected from FRT, attP, attB, ITR and variants thereof, herein, the first RRS is capable of pairing with the third RRS. ### Claim 22 (depends on 13) The method of claim 13, wherein the second RRS is one selected from FRT, attP, attB, ITR and variants thereof, wherein the fourth RRS is one selected from FRT, attP, attB, ITR and variants thereof, herein, the second RRS is capable of pairing with the fourth RRS. ### Claim 23 (depends on 21) The method of claim 21, wherein the SSR is one selected from a flippase (FLP), an integrase and a transposase. ### Claim 24 (depends on 13) The method of claim 13, wherein the cell including one or more artificial recombinant chromosome can undergo mitosis or meiosis. ### Claim 25 (depends on 13) The method of claim 13, wherein the cell including one or more artificial recombinant chromosome expresses a protein encoded by a gene present in the second fragment. ### Claim 26 (canceled) ### Claim 27 A method for making a transgenic non-human animal using a cell including one or more artificial recombinant chromosome, the method comprising i) preparing a first targeted cell and a second target cell, wherein the first targeted cell is an embryonic stem cell, wherein the first targeted cell comprises a first targeted chromosome, and the second targeted cell comprises a second targeted chromosome, wherein the first targeted chromosome includes a first part, a first recombinase recognition sequence (a first RRS), a first fragment, a second recombinase recognition sequence (a second RRS) and a second part, herein, the first part includes one of telomere ends located on both sides of the first targeted chromosome, and the second part includes the other one of telomere ends located on both sides of the first targeted chromosome, herein, the first fragment is located between the first RRS and the second RRS, wherein the second targeted chromosome includes a third part, a third recombinase recognition sequence (a third RRS), a second fragment, a fourth recombinase recognition sequence (a fourth RRS) and a fourth part, herein, the third part includes one of telomere ends located on both sides of the second targeted chromosome, and the fourth part includes the other one of the telomere ends located on both sides of the second targeted chromosome, herein, the second fragment is located between the third RRS and the fourth RRS; ii) producing one or more microcells using the second targeted cell, wherein the one or more microcells comprises the second targeted chromosome or a fragment thereof, herein, the fragment of the second targeted chromosome includes the third RRS, the second fragment and the fourth RRS; iii) producing a fusion cell using the first targeted cell and the one or more microcells, wherein the fusion cell comprises the first targeted chromosome and the second targeted chromosome, or the first targeted chromosome and the fragment of the second targeted chromosome; iv) producing a cell including an artificial recombinant chromosome by treating the fusion cell with a site specific recombinase (SSR), wherein the SSR induces a recombination by recognizing a pairing of the first RRS present in the first targeted chromosome and the third RRS present in the second targeted chromosome, and a pairing of the second RRS present in the first targeted chromosome and the fourth RRS present in the second targeted chromosome, herein, the first fragment present in the first targeted chromosome is exchanged for the second fragment present in the second targeted chromosome, thereby, a first artificial recombinant chromosome including the first part, the second fragment and the second part is produced; and v) implanting a chimeric blastocyst comprising the first artificial recombinant chromosome in a surrogate mother's uterus to produce an offspring, herein, the chimeric blastocyst is produced by injecting the cell including the first artificial recombinant chromosome into a blastocyst.