Endocytic pathogens such as protein aggregates, viruses, protein toxins, and bacteria have evolved remarkable ways to enter the cell, disrupt homeostasis, and cause cell death. We use these agents both as probes to understand normal cellular trafficking and signaling events, and to find key targets for therapy.
Cells have elaborate mechanisms of sensing diverse stresses (oxidative damage, nutrient deprivation, DNA breaks, etc), and must either repair damage or induce cell death. Misregulation of these pathways results in diseases such as cancer and Alzheimer’s. We would like to understand how these signals connect to the death pathway in health and disease in order to improve therapies.
Much of the work we do utilizes genetic screens enabled by novel high-coverage shRNA libraries (~25 shRNAs/gene) we have developed. The high coverage greatly reduces false positive and false negative results that have plagued traditional RNAi strategies. We use a pooled format that can be rapidly screened in large bioreactors, and analyze screens by deep sequencing to quantify changes in shRNA abundance.
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