Transgenomic Mitochondria, Transmitochondrial Cells and Organisms, and Methods of Making and Using

Abstract

This invention provides transgenomic mitochondria, transmitochondrial cells and organisms, and the materials and methods for making such mitochondria, cells and organisms.

1 . A method for making a transgenomic mitochondrion incorporating an exogenous mitochondrial genome comprising the steps of;
 a) isolating a mitochondrial genome from a eukaryotic cell to yield an exogenous mitochondrial genome;   b) modifying said isolated exogenous mitochondrial genome by insertion of one or more selectable markers, screening markers, replication sequences, or combinations thereof;   c) introducing said modified isolated exogenous mitochondrial genome into a host mitochondrion from a second eukaryotic cell;   d) removing said one or more selectable markers, replication sequences, or combinations thereof, from said modified isolated exogenous mitochondrial genome within said host mitochondrion; and   e) isolating said transgenomic mitochondrion from said second eukaryotic cell.   
     
     
         2 . The method of  claim 1 , wherein said host mitochondrion from said second eukaryotic cell is a yeast mitochondrion. 
     
     
         3 . The method of  claim 2 , wherein said yeast mitochondrion is substantially free of endogenous mitochondrial DNA. 
     
     
         4 . The method of  claim 1 , wherein said exogenous mitochondrial genome is an isolated animal mitochondrial genome. 
     
     
         5 . The method of  claim 4 , wherein said isolated animal mitochondrial genome is modified prior to introduction into said isolated yeast mitochondrion. 
     
     
         6 . The method of  claim 5 , wherein said isolated animal mitochondrial genome is modified by one or more selectable markers, screening markers, replication sequences, or combinations thereof, said selectable markers, screening markers, and replication sequences having a prokaryotic origin. 
     
     
         7 . The method of  claim 5 , wherein said isolated animal mitochondrial genome is modified by one or more selectable markers, screening markers, replication sequences, or combinations thereof, said selectable markers, screening markers, and replication sequences having a eukaryotic origin. 
     
     
         8 . The method of  claim 5 , wherein said isolated animal mitochondrial genome is modified by one or more selectable markers, screening markers, replication sequences, or combinations thereof, said selectable markers, screening markers, and replication sequences having a synthetic origin. 
     
     
         9 . The method of  claim 5 , wherein said isolated animal mitochondrial genome constitutes at least 80% of naturally occurring said isolated animal mitochondrial genome. 
     
     
         10 . The method of  claim 9 , wherein said isolated animal mitochondrial genome is introduced into said host yeast mitochondrion. 
     
     
         11 . The method of  claim 10 , wherein said introduction of said isolated animal mitochondrial genome into said host yeast mitochondrion is by particle bombardment. 
     
     
         12 . A transgenomic mitochondrion comprising:
 a mitochondrion having an outer membrane and an inner membrane;   a matrix located inside said inner membrane; and   an exogenous mitochondrial genome that is incorporated into and stably replicating within said matrix of said mitochondrion.   
     
     
         13 . The transgenomic mitochondrion of  claim 12 , wherein said mitochondrion is an isolated yeast mitochondrion comprising an exogenous mitochondrial genome. 
     
     
         14 . The transgenomic mitochondrion of  claim 12 , wherein said exogenous mitochondrial genome comprises an exogenous animal mitochondrial genome. 
     
     
         15 . The transgenomic mitochondrion of  claim 14 , wherein said animal mitochondrial genome is selected from the group consisting of avian, mammalian, murine, bovine, porcine, or human mitochondrial genomes. 
     
     
         16 . The transgenomic mitochondrion of  claim 13 , wherein said mitochondrion is a yeast mitochondrion comprising an animal mitochondrial genome. 
     
     
         17 . The transgenomic mitochondrion of  claim 15 , wherein said animal mitochondrial genome is a mouse mitochondrial genome. 
     
     
         18 . The transgenomic mitochondrion of  claim 15 , wherein said animal mitochondrial genome is a human mitochondrial genome. 
     
     
         19 . The transgenomic mitochondrion of  claim 13 , wherein said isolated yeast mitochondrion is substantially free of endogenous yeast mitochondrial DNA. 
     
     
         20 . A transmitochondrial cell comprising:
 a eukaryotic cell having a plasma membrane, cytoplasm, a nucleus; and   a transgenomic mitochondrion incorporated into said cytoplasm of said eukaryotic cell, said transgenomic mitochondrion having a modified exogenous mitochondrial genome within said mitochondrion.   
     
     
         21 . The transmitochondrial cell of  claim 20 , wherein said transgenomic mitochondrion is not endogenous to said eukaryotic cell. 
     
     
         22 . The transmitochondrial cell of  claim 20 , wherein said modified exogenous mitochondrial genome is stably replicating within said transgenomic mitochondrion within said eukaryotic cell. 
     
     
         23 . The transmitochondrial cell of  claim 21 , wherein said transgenomic mitochondrion is a yeast mitochondrion. 
     
     
         24 . The transmitochondrial cell of  claim 22 , wherein said modified exogenous mitochondrial genome is an animal genome.