This technology is an in vivo system in which a covalent bond can be formed between any two proteins in a cell-type specific manner for the purpose of immunoprecipitation, visualization, or modifying the activity of a target protein.
Current methods to fuse proteins to other proteins or DNA for immunoprecipitation or visualization are not cell-type specific. Whole tissue chromatin immunoprecipitation (ChIP), for example, is not able to characterize protein-DNA binding in specific cell types within complex tissues. Using a tagging system to covalently bind two proteins of interest in vivo is also not cell-type specific and may require large antibodies. There are currently no assay platforms available that enable direct covalent bonding of two peptides in vivo with cell type-specificity without interfering with native protein structure, localization, or function.
This assay is based on the already existing SpyTag-SpyCatcher system whereby a peptide tag forms a rapid covalent bond to a protein, through engineering a bacterial adhesin. The effector protein can be an epitope, fluorescent protein, or enzyme. The “SpyCatcher::effector” protein is only expressed in the cell type of interest. By co-expressing the SpyTagged protein and the SpyCatcher-tagged effector in a specific cell type, the two tags form a covalent bond in vivo, fusing the two proteins.
This technology has been validated in Drosophila melanogaster tissues.
IR CU22274
Licensing Contact: Jerry Kokoshka
