Our lab is interested in building technologies that help answer fundamental biological questions. Below you'll see a few of our technologies and their applications.
Current methods of generating cell lines with defined alterations are slow, costly, and not easily scaled to address the hundreds of millions of variants of unknown function. Our team has developed a strategy to generate thousands of yeast mutants, each with a defined genetic alteration (point mutation, deletion, or insertion) in a single reaction. We have also generated a pipeline for simultaneously tracking the fitness of all of the resulting mutants en masse. We are applying our technology to gain insights into the coding and non-coding portions of the genome, optimize metabolic pathways and better understand how point mutations influence protein structure and function.
We created a second generation CRISPR-activator, which allows for potent gene activation across a variety of model organisms (human cells, mice, flies and yeast). We are currently using our activator for genome-wide screens and to study factors involved in cellular differentiation.
Synthetic gene drives
Gene drives are selfish genetic elements that "cheat" Mendelian inheritance. We were among the first to generate synthetic gene drives using Cas9. Our team is using this tool to perform rapid combinatorial knockouts within the clinically relevant fungal pathogen Candida albicans, along with exploring other exciting applications within yeast.
Tuned guide RNAs
We developed a system to endow Cas9 with single nucleotide specificity, through the development of tuned guide RNAs. With this method in hand we developed a "genomic surveillance system" which prevents cells from gaining undesired mutations.
DNA barcoded libraries
By employing DNA barcodes, we are able to track the growth of dozens of cellular models at a time. We use these pools of barcoded cells to perform multiplex screens aimed at understanding the genes involved in tolerance to misfolded proteins.
We uncovered that Cas9 nuclease activity can be regulated by altering the length of the complexed gRNA. Using this knowledge we were able to perform simultaneous genome editing, gene activation and gene repression using a single Cas9 protein (Cas9-VPR). We plan to use this discovery to dissect the complex interplay between factors involved in adaptation to stress and drug resistance.