# US 8,173,861 B2 · Transgenic pig model for a hereditary neurodegenerative autosomal dominant disease > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 8,173,861 B2 - Document ID: US8173861B2 - Status: Granted - Kind: B2 - Application number: 12/298,974 - Filed: 2007-04-30 - Published: 2009-12-10 - Granted: 2012-05-08 - Earliest claimed: 2006-05-01 - Assignees: Aarhus Universitet - Inventors: Lone Bruhn Madsen; Christian Bendixen; Knud Larsen; Connie Jakobsen Juhl; Bo Thomsen - Web page: https://patentgrep.com/patent/US8173861B2 ## CPC - G01N 33/5088: {of vertebrates} - A01K 67/0275: Genetically modified vertebrates, e.g. transgenic - A61K 49/0008: {Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure} - C07K 14/4711: {Alzheimer's disease; Amyloid plaque core protein} - C12N 15/8509: {for producing genetically modified animals, e.g. transgenic} - C12N 15/8778: {Swine embryos} - A01K 2217/00: Genetically modified animals - A01K 2217/056: due to mutation of coding region of the transgene (dominant negative) - A01K 2227/108: Swine - A01K 2267/03: Animal model, e.g. for test or diseases - A01K 2267/0306: Animal model for genetic diseases - A01K 2267/0312: Animal model for Alzheimer's disease - A01K 2267/0318: Animal model for neurodegenerative disease, e.g. non- Alzheimer's - A01K 2267/035: Animal model for multifactorial diseases - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - A01K: ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS - 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) - 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 discloses a non-human animal model for a hereditary autosomal dominant disease. The non-human animal model expresses at least one phenotype associated with the disease and is obtained by a genetic determinant. The invention also relates to sperm cells and embryos comprising the genetic determinant for an autosomal dominant disease. Furthermore, methods for producing the non-human animal model, sperm cell, and embryos are disclosed. ## Claims ### Claim 1 A transgenic pig model for a hereditary neurodegenerative autosomal dominant disease, comprising a transgenic pig whose genome comprises a DNA sequence encoding a protein involved in the development of a hereditary neurodeqenerative autosomal dominant disease, wherein the transgenic pig expresses the protein in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue. ### Claim 2 (depends on 1) The pig model according to claim 1, wherein said hereditary neurodegenerative autosomal dominant disease is amyotrophic lateral sclerosis. ### Claim 3 (depends on 1) The pig model according to claim 1, wherein said hereditary neurodegenerative autosomal dominant disease is Alzheimer's Disease. ### Claim 4 (depends on 1) The pig model according to claim 1, wherein said hereditary neurodegenerative autosomal dominant disease is Parkinson's Disease. ### Claim 5 (depends on 1) The pig model according to claim 1, wherein said hereditary neurodegenerative autosomal dominant disease is a disease related to trinucleotide repeats. ### Claim 6 (depends on 1) The pig model according to claim 1, wherein said hereditary neurodegenerative autosomal dominant disease is Huntington's chorea. ### Claim 7 (depends on 1) The pig model according to claim 1, wherein said hereditary neurodegenerative autosomal dominant disease is dyschondroplasia. ### Claim 8 (depends on 1) The transgenic pig model according to claim 1 obtainable by a sperm mediated gene transfer method (SMGT) comprising the steps of: i) providing semen from a male pig, ii) providing a DNA sequence encoding a protein involved in the development of a hereditary neurodeqenerative autosomal dominant disease, wherein the transgenic pig expresses the protein in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue when the protein is expressed in said pig model, iii) contacting said semen and said genetic material, iv) fertilising an oocyte from a female pig with the semen and the genetic determinant, and v) incubating said fertilised oocyte under conditions allowing said fertilised oocyte to develop into said pig model. ### Claim 9 A method for producing a transgenic pig model for a hereditary neurodegenerative autosomal dominant disease, comprising a transgenic pig whose genome comprises a DNA sequence encoding a protein involved in the development of a hereditary neurodeqenerative autosomal dominant disease, wherein the transgenic pig expresses the protein in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue, said method comprising the steps of: i) providing semen from a male pig, ii) providing a DNA sequence encoding a protein involved in the development of a hereditary neurodegenerative autosomal dominant disease, iii) contacting said semen and said DNA sequence, iv) fertilising an oocyte from a female pig with the semen and the genetic material, and v) incubating said fertilised oocyte under conditions allowing said fertilised oocyte to develop into said transgenic pig, wherein the transgenic pig expresses the protein in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue. ### Claim 10 (depends on 9) The method of claim 9, wherein the step of contacting said semen and said genetic material occurs in a buffer composition comprising (a) glucose and (b) a citrate salt and/or a bicarbonate salt, optionally in combination with (c) a compound capable of chelating divalent metal ions, and further optionally in combination with (d) Bovine Serum Albumin (BSA), including any combination of (a) and (b) with (c) or (d). ### Claim 11 A method for evaluating the response of a therapeutical treatment of a hereditary disease, said method comprising the steps of: a. providing a transgenic pig model for a hereditary neurodegenerative autosomal dominant disease, comprising a transgenic pig whose genome comprises a DNA sequence encoding a protein involved in the development of a hereditary neurodeqenerative autosomal dominant disease, wherein the transgenic pig expresses the protein in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue, b. treating said pig with at least one pharmaceutical composition exerting an effect on the reduction in the number of neurons in the nerve tissue or the accumulation of protein aggregates in the nerve tissue, and c. evaluating the effect observed. ### Claim 12 (depends on 11) The method of claim 11 comprising the further step of advising on medical treatment based on the afore-mentioned observed effects. ### Claim 13 A transgenic pig sperm cell comprising a DNA sequence encoding a protein involved in the development of a hereditary neurodegenerative autosomal dominant disease wherein said protein is expressed in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue when expressed in a pig host organism. ### Claim 14 (depends on 13) The pig sperm cell according to claim 13, wherein the DNA sequence is of mammalian origin, including human origin or porcine origin. ### Claim 15 A method for producing a transgenic pig sperm cell comprising a DNA sequence encoding a protein involved in the development of a hereditary neurodegenerative autosomal dominant disease, wherein said protein is expressed in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue when expressed in a pig host organism, said method comprising the steps of: a. providing a pig sperm cell, b. providing a DNA sequence encoding a protein involved in the development of a hereditary neurodeqenerative autosomal dominant disease wherein said protein is expressed in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue when expressed in a pig host organism, and c. contacting said pig sperm cell and said DNA sequence, wherein said contacting results in the uptake of the DNA sequence into the pig sperm cell. ### Claim 16 (depends on 13) A method for fertilising an oocyte by sperm-mediated gene transfer, said method comprising the steps of providing a pig sperm cell according to claim 13, and introducing said pig sperm cell into the oocyte to be fertilised, wherein said disease is a protein conformation disease. ### Claim 17 A method for fertilising an oocyte by sperm-mediated gene transfer, said method comprising the step of providing a composition comprising a pig sperm cell in combination with a DNA sequence encoding a protein involved in the development of an hereditary neurodegenerative autosomal dominant disease wherein said protein is expressed in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue when expressed in a pig host organism. ### Claim 18 A transgenic embryo obtained by fertilising an oocyte with a pig sperm cell comprising a DNA sequence encoding a protein involved in the development of a hereditary neurodegenerative autosomal dominant disease wherein said protein is expressed in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue when expressed in a pig host organism. ### Claim 19 A transgenic embryo obtained by fertilising an oocyte with a composition comprising a DNA sequence encoding a protein involved in the development of a hereditary neurodegenerative autosomal dominant disease, wherein said protein is expressed in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue when expressed in a pig host organism. ### Claim 20 A method for the cultivation and development of a transgenic embryo obtained by fertilising an oocyte with a pig sperm cell comprising a DNA sequence encoding a protein involved in the development of a hereditary neurodegenerative autosomal dominant disease, said method comprising the step of cultivating said embryo under conditions allowing the embryo to develop into a pig offspring expressing said protein in nerve tissue and exhibiting a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue. ### Claim 21 A method for screening the efficacy of a pharmaceutical composition, said method comprising the steps of: a. providing a transgenic pig model for a hereditary neurodegenerative autosomal dominant disease, comprising a transgenic pig whose genome comprises a DNA sequence encoding a protein involved in the development of a hereditary neurodegenerative autosomal dominant disease, wherein the transgenic pig expresses the protein in nerve tissue and exhibits a reduction in the number of neurons in the nerve tissue and an accumulation of protein aggregates in the nerve tissue, b. expressing in said pig model said DNA sequence involved in the development of a hereditary neurodegenerative autosomal dominant disease, c. administering to said pig the pharmaceutical composition, the efficacy of which is to be evaluated, and d. evaluating the effect, if any, of the pharmaceutical composition on the reduction in the number of neurons in the nerve tissue or the accumulation of protein aggregates in the nerve tissue exerted by the DNA sequence when expressed in the pig model. ### Claim 22 (depends on 1) The pig model according to claim 1, wherein said DNA sequence is expressed in a transient manner.