SBIR Phase I: Engineered large-scale genomic deletions with a universal targeted cleavage system

This Small Business Innovation Research (SBIR) Phase I project seeks to develop a technology for specifically removing large regions of genomic DNA from culture cells. Genome editing technologies have transformed cell biology by enabling the targeted knock-out or replacement of genomic loci. These edits permit the introduction of disease-relevant mutations in cellular and animal models for basic research and drug discovery applications. Current technologies rely on custom enzymes to cleave specific genomic sites, and are therefore limited in their ability to cleave multiple genomic sites in parallel. Cas9 is a recently discovered bacterial protein that is an RNA-guided DNA endonuclease. The company will test the ability of Cas9 to cleave mouse genomic DNA in two locations simultaneously and excise the intervening sequence from the genome. Both short and long genomic loci will be removed by targeting Cas9 to nearby and distant loci, respectively. The broader impact/commercial potential of this project, if successful, will be the ability to generate cellular and animal models of genetic diseases caused by large-scale genomic deletions and rearrangements. Successful demonstration of the ability to excise genomic loci in cultured cells will permit the development of this technology for excising regions of mouse embryo genomic DNA and the generation of new animal disease models. Large scale deletions can remove introns, exons, intergenic regions, or even entire gene clusters, recapitulating causative mutations of inherited disorders, a need poorly met by currently available genome engineering technologies. The cellular and animal models developed using this technology will further basic research and the understanding of how particular genetic loci control biological pathways and will facilitate the identification and development of new drug targets.