The Genomic Revolution has clearly demonstrated the importance of genome sequence and structure in defining the disease state of cells and tissues, as well as the response to regenerative stimuli. However, there is currently no general method to efficiently and specifically alter targeted gene sequences within mammalian chromosomes. Therefore we are developing a broadly applicable technology for editing endogenous gene sequences and regulating the expression of gene products. Over the past decade, our laboratory has pioneered the development of synthetic DNA-binding proteins based on the canonical zinc finger peptide motif. These studies have established a family of programmable transcription factors that can be targeted to a distinct site within the human genome with high affinity and specificity. More recently, we have combined the modular DNA-binding specificity of artificial zinc finger proteins with the potent catalytic activity of recombinase domains that have been engineered through directed protein evolution. These fusion proteins are capable of targeting a specific recognition site within the human genome and coordinating the efficient excision of endogenous gene sequences, as well as the integration of transfected plasmid DNA into a defined chromosomal site. This technology represents an exciting approach to tailoring genome sequences for basic biological studies and controlling cell activity in the context of tissue regeneration and disease treatment and prevention.

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