B-MoGen Biotechnologies Inc — Department of Health and Human Services STTR Phase I: OD

Description

Project Abstract Mitochondria are double membrane bound organelles that are found in all eukaryotic cells and carry a specialized circularkb genomeThere is a group of devastating human genetic diseases caused by inherited or spontaneous mutation of genes encoded within the mitochondrial genomeThese diseases can cause a wide array of medical problems from subtle muscular weakness to extreme neuromuscular problems such as loss of balance and coordinationseizuresstrokedementia and deathFurtherit is estimated thatinchildren are diagnosed with a mitochondrial disease each yearDespite the importance of the mitochondrial genome in maintaining normal cellular functionas well as its role in promoting mitochondrial dysfunction when mutatedno method exists to alter the genome of mitochondria in a site specific manneruntil nowWe have recently developed the first reliable method to introduce specific mutations into the mitochondrial genome of vertebrate animal cellsWe have found that two pairs of site specific DNA binding transcription activator likeTALdomain endo nickasesmitoTALE nicaksescan be directed to the mitochondria and that their activity results in generation of an intact mitochondrial genomebut with a deletion between the sites of the mtDNA nicksUsing a second pair of mitochondrial targeted TAL endonucleasesmitoTALENsto make a double strand DNA break in between the nick siteswe can induce the selective loss of wild type mitochondrial genomes which did not undergo the deletionThuswe can seed mtDNA deletions using mitoTALE nickases and enrich for edited mtDNA genomes using mitoTALENsWe have efficiently induced mtDNA deletions in zebrafish embryos and human and mouse cell linesIn this proposalwe will extend the innovations in mammalian cells by engineering targeted and precise mtDNA deletions systemically in miceWe will also develop inducible systems for expressing the mitoTALENswhich means we will make it possible to createconditionalloss of function of mitochondrial genes in miceThis inducible mouse model system could be highly useful in settings in which the mitochondrial deletion would cause a selective disadvantage to such cells during early mouse developmentWe will create mice carrying an inducible deletion that causes Kearns Sayre syndromeKSSin humansone of the most common mitochondrial genetic diseaseThe production of these novel mouse models will be useful for academic and pharmaceutical companies to test novel regenerative stem cell technologies Project Narrative Although there have been significant advancements in gene editing over the past two decadesnone have been able to precisely edit the mitochondrial genomeuntil nowThe goal of the proposed project is to use mitochondrial gene editing technology to generate an inducible mouse model of mitochondrial genetic diseaseThis project will result in a set of mouse models necessary to identify and test novel therapeutics for the treatment of mitochondrial disease