Genomically rewritten and tailored humanized mouse models for various organ disorders

Summary This proposal leverages Big DNA technology to produce mouse models of a variety of human diseases. The underlying hypothesis tested in our earlier work and continued in this proposal is that changes in human non- coding DNA that underlie human disease susceptibility will translate into appropriate disease phenotypes in the mouse when the entire human genic region of interest is inserted into a predefined location in the mouse genome. We refer to such animals as GREAT-GEMMs (Genomically Rewritten and Tailored Genetically engineered moused models). Furthermore, GREAT-GEMMs will serve as a resource for testing therapeutic interventions against human disease. These animals will be uniquely useful for cutting edge therapies like gene therapies, CRISPR based therapies and allele specific oligonucleotide (ASO) therapies which depend on exact human DNA sequences as targets. Our work is based on our recently developed ability to synthesize and precisely edit 50-300+ kilobase DNA constructs, and then to precisely delivery them to mouse embryonic stem cells (mESCs) and subsequently produce mouse models for human disease.