Department of Molecular and Cell Biology
University of California, Berkeley
We are using zinc finger nucleases and TALENs to edit the genomes of human iPS cells to engineer gene fusions that express at endogenous levels membrane trafficking and cytoskeletal proteins fused to fluorescent proteins. These engineered iPS cells offer significant advantages for cell biology studies over traditionally used tissue culture cells. The genome editing insures that protein stoichiometry is preserved, the cells are true diploids lacking the chromosome duplications and translocations that characterize standard tissue culture cell lines, and iPS cells are derived from normal tissues rather than from tumors, which are the source of most tissue culture cell lines. Moreover, because the iPS cells can be differentiated to many different cell types, we are able to test how differentiation affects basic biological processes. In total, we are endeavoring to set a new standard for studies of human cell biology with the goal of attaining a more complete and accurate understanding of basic human cell biology.