Mitochondrial-nuclear exchanged cells, tissues, organs and animals

Abstract

Provided herein are mitochondrial-nuclear exchanged cells and animals comprising mitochondrial DNA (mtDNA) from one subject and nuclear DNA (nDNA) from a different subject. Methods for producing a mitochondrial-nuclear exchanged animal and animals made by the methods are provided. Also provided are methods of screening for agents useful for treating a disease or disorder using mitochondrial-nuclear exchanged animals or cells, tissues or organs thereof.

1 . A cell comprising mitochondrial DNA (mtDNA) from a subject susceptible to a disease or disorder and nuclear DNA (NDNA) from a subject that is not susceptible to the disease or disorder. 
   
   
       2 . The cell of  claim 1 , wherein the nDNA is from a wild-type subject. 
   
   
       3 . The cell of  claim 1 , wherein the nDNA is from a subject resistant to the disease or disorder. 
   
   
       4 . The cell of  claim 1 , wherein the disease or disorder is selected from the group consisting of cancer, cardiovascular disease, diabetes, neurological disorder, aging, metabolic disorder, immune disorder, obesity, and musculoskeletal disorder. 
   
   
       5 . The cell of  claim 1 , wherein the cell is an oocyte. 
   
   
       6 . The cell of  claim 1 , wherein the cell is an embryonic cell. 
   
   
       7 . A zygote or an embryo comprising the cell of  claim 1 . 
   
   
       8 . The embryo of  claim 7 , wherein the embryo is a pro-nuclear embryo. 
   
   
       9 . A chimeric animal comprising a plurality of cells of  claim 1 . 
   
   
       10 . The chimeric animal of  claim 9 , wherein the animal further comprises a mutation in a gene associated with the disease or disorder. 
   
   
       11 . The chimeric animal of  claim 10 , wherein the gene is not expressed or the protein expressed by the gene is non-functional. 
   
   
       12 . The chimeric animal of  claim 9 , wherein the animal is a mouse comprising mtDNA from C57BL/6J mice and nDNA from C3H/HeN mice. 
   
   
       13 . The chimeric animal of  claim 9 , wherein the animal is a female. 
   
   
       14 . A progeny animal of the animal of  claim 9 . 
   
   
       15 . A progeny animal resulting from a cross between the chimeric animal of  claim 13  and a knockout mouse, wherein the knockout mouse comprises a mutation in at least one gene associated with the disease or disorder such that the gene is not expressed or the protein expressed by the gene is not functional. 
   
   
       16 . The progeny animal of  claim 15 , wherein the knockout mouse is susceptible to the disease or disorder. 
   
   
       17 . A method of screening for agents useful for treating a disease or disorder comprising:
 (a) administering to the animal of  claim 9  an agent to be tested; and   (b) determining whether the agent prevents or reduces one or more symptoms of the disease or disorder.   
   
   
       18 . A cell comprising mitochondrial DNA (mtDNA) from a subject resistant to a disease or disorder and nuclear DNA (nDNA) from a wild-type subject or a subject that is susceptible to the disease or disorder. 
   
   
       19 . The cell of  claim 18 , wherein the NDNA is from a wild-type subject. 
   
   
       20 . The cell of  claim 18 , wherein the NDNA is from a subject susceptible to the disease or disorder. 
   
   
       21 . The cell of  claim 18 , wherein the disease or disorder is selected from the group consisting of cancer, cardiovascular disease, diabetes, neurological disorder, aging, metabolic disorder, immune disorder, obesity, and musculoskeletal disorder. 
   
   
       22 . The cell of  claim 18 , wherein the cell is an oocyte. 
   
   
       23 . The cell of  claim 18 , wherein the cell is an embryonic cell. 
   
   
       24 . A zygote or an embryo comprising the cell of  claim 18 . 
   
   
       25 . The embryo of  claim 24 , wherein the embryo is a pro-nuclear embryo. 
   
   
       26 . A chimeric animal comprising a plurality of cells of  claim 18 . 
   
   
       27 . The chimeric animal of  claim 26 , wherein the animal is a mouse comprising mtDNA from NZB/B1NJ mice and nDNA from FVB/N-TgN(MMTVPyMT) mice. 
   
   
       28 . A method for producing a mitochondrial-nuclear exchanged animal comprising:
 (a) selecting an animal susceptible to a disease or disorder;   (b) selecting an animal that is not susceptible to the disease or disorder;   (c) harvesting a pro-nuclear embryo from each of the animals of steps (a) and (b);   (d) enucleating the embryos of step (c);   (e) transferring the nucleus from the embryo of the animal that is not susceptible to the disease or disorder to the enucleated embryo of the animal susceptible to the disease or disorder to make a resulting embryo, wherein the resulting embryo has mtDNA from the animal susceptible to the disease or disorder and the nDNA from the animal that is not susceptible to the disease or disorder; and   (f) transferring the resulting embryo into an appropriate host and allowing the transferred embryo to develop into a progeny animal, wherein the progeny animal is a mitochondrial-nuclear exchanged animal.   
   
   
       29 . The method of  claim 28 , wherein the animal that is not susceptible to the disease or disorder is from a wild-type animal or an animal resistant to the disease or disorder. 
   
   
       30 . The method of  claim 28 , further comprising selecting a female mitochondrial-nuclear exchanged animal. 
   
   
       31 . A method of generating progeny of the female mitochondrial-nuclear exchanged animal of  claim 30  by crossing the female mitochondrial exchanged animal with a knockout animal, wherein the knockout animal comprises a mutation in at least one gene associated with the disease or disorder such that the gene is not expressed or the protein expressed by the gene is not functional. 
   
   
       32 . The method of  claim 31 , further comprising selecting progeny animals of the cross that comprise mtDNA from the mitochondrial-nuclear exchanged animals and nDNA from the knockout mouse. 
   
   
       33 . The method of  claim 28 , wherein the disease or disorder is selected from the group consisting of cancer, cardiovascular disease, diabetes, neurological disorder, aging, metabolic disorder, immune disorder, obesity, and musculoskeletal disorder. 
   
   
       34 . A mitochondrial-nuclear exchanged animal made by the method of  claim 28 . 
   
   
       35 . A method of screening for agents useful for treating a disease or disorder comprising the steps of:
 (a) providing a mitochondrial-nuclear exchanged animal comprising mtDNA from an animal susceptible to the disease or disorder and nDNA from an animal not susceptible to the disease or disorder;   (b) administering to the animal an agent to be tested; and   (c) determining whether the agent prevents or reduces one or more symptoms of the disease or disorder.   
   
   
       36 . The method of  claim 35 , wherein the animal not susceptible to the disease or disorder is a wild-type animal or an animal resistant to the disease or disorder. 
   
   
       37 . The method of  claim 35 , wherein the disease or disorder is selected from the group consisting of cancer, cardiovascular disease, diabetes, neurological disorder, aging, metabolic disorder, immune disorder, obesity, and musculoskeletal disorder. 
   
   
       38 . The method of  claim 37 , wherein the disease is cancer and wherein the mitochondrial-nuclear exchanged animal is a mouse comprising mtDNA from a mouse selected from the group consisting of FVB/N-TgN(MMTVPyMT), AKR/J, and A/J mice. 
   
   
       39 . The method of  claim 37 , wherein the disease is cardiovascular disease and wherein the mitochondrial-nuclear exchanged animal is a mouse comprising mtDNA from C57BL/6J or DBA/2J mice. 
   
   
       40 . The method of  claim 35 , wherein the mitochondrial-nuclear exchanged animal is a mouse comprising mtDNA from C57BL/6J mice and NDNA from C3H/HeN mice. 
   
   
       41 . The method of  claim 35 , wherein the mitochondrial-nuclear exchanged animal is a mouse comprising NDNA from a mouse selected from the group consisting of C57BL/6J, 129, A/J, BALB/c or C3H/HeN mice. 
   
   
       42 . A method of screening for agents useful for treating a disease or disorder comprising the steps of:
 (a) providing a mitochondrial-nuclear exchanged cell, tissue or organ comprising mtDNA from an animal susceptible to the disease or disorder and nDNA from an animal not susceptible to the disease or disorder;   (b) contacting the cell, tissue or organ with an agent to be tested; and   (c) determining whether the agent prevents or reduces one or more symptoms of the disease or disorder.   
   
   
       43 . The method of  claim 42 , wherein the cells, tissues or organs are obtained from a mitochondrial-nuclear exchanged animal or progeny thereof. 
   
   
       44 . The method of  claim 42 , wherein the animal not susceptible to the disease or disorder is a wild-type animal or an animal resistant to the disease or disorder. 
   
   
       45 . The method of  claim 42 , wherein the disease or disorder is selected from the group consisting of cancer, cardiovascular disease, diabetes, neurological disorder, aging, metabolic disorder, immune disorder, obesity, and musculoskeletal disorder. 
   
   
       46 . The method of  claim 43 , wherein the disease is cancer and wherein the mitochondrial-nuclear exchanged animal is a mouse comprising mtDNA from a mouse selected from the group consisting of FVB/N-TgN(MMTVPyMT), AKR/J, and A/J mice. 
   
   
       47 . The method of  claim 43 , wherein the disease is cardiovascular disease and wherein the mitochondrial-nuclear exchanged animal is a mouse comprising mtDNA from C57BL/6J or DBA/2J mice. 
   
   
       48 . The method of  claim 43 , wherein the mitochondrial-nuclear exchanged animal is a mouse comprising mtDNA from C57BL/6J mice and nDNA from C3H/HeN mice. 
   
   
       49 . The method of  claim 43 , wherein the mitochondrial-nuclear exchanged animal is a mouse comprising nDNA from a mouse selected from the group consisting of C57BL/6J, 129, A/J, BALB/c or C3H/HeN mice.