BBMB Research Seminars

October 25, 2007
David Segal
Genome Center/Pharmacology
University of California, Davis

"Editing and imaging genomes using engineered zinc finger proteins”
1414 Molecular Biology Buidling
4:10 p.m.

Abstract:

Zinc fingers are a type of DNA-binding domain used by many transcription factors. Our group is actively developing technology for engineering custom zinc finger proteins. These proteins have been reprogrammed to bind to DNA sequences of therapeutic or diagnostic interest. We anticipate the direct detection of double-stranded DNA may have advantages for the detection of specific DNA sequence information in live cell imaging, field kits, or molecular diagnostics. One imaging applications we have developed is SEquence-Enabled Reassembly (SEER), in which the presence of an appropriate DNA target generates a fluorescent signal. Other types of functional domains can be attached to engineered zinc finger proteins various to create site-specific tools and potential therapeutics. Nucleases appended to zinc fingers have been used to stimulate highly-efficient homologous recombination at specific endogenous loci. Using this approach, targeted genomic modifications (insertions or substitutions) have been observed to occur in 10-20% of treated cells in the absence of any selection. This approach could therefore enable transgenic and “knock-out” type experiments in animals which traditionally have not been as genetically accessible as mice. We are using this approach to develop gene therapies.