DEMONSTRATING COMMERCIAL POTENTIAL OF ZINC FINGER PROTEINS FOR GENERATING VALUE-ADDED CROPS

Not Available Zinc finger proteins (ZFPs) of the C2-H2 class are DNA-binding proteins that are ubiquitous in nature. By modifying the amino acids of the recognition helices according to a recently devised recognition code, ZFP's can be designed to bind DNA sequences of choice. This development has made possible the construction of chimeric transcription repressors and activators for regulating genes in plants. We propose to use this emerging technology to alter fatty acid desaturation in both a model system and commercially relevant crop species. In Phase I, we successfully constructed ZFPs capable of recognizing target sequences in a fatty acid desaturase gene of soybean (Glycine max). The custom DNA binding proteins were produced in E. coli and shown to bind to their corresponding target sequences with nanomolar affinities. In Phase II, a second set of ZFPs will be designed for the Arabidopsis version of the desaturase gene. The ZFPs will be optimized for enhanced binding affinity and specificity, they will be inserted into plant expression vectors, and the resulting constructs will be tested in cell culture and whole plants. Success will constitute proof-of-principle for the use of designed ZFPs as a method to alter the expression of genes in plants