I will discuss three novel technologies that our laboratory has developed in recent years. In the first part of the talk, I will describe our work on engineering variants of the RNA-guided endonuclease, Cas9, for the selective control of gene expression. I will then share our method for endowing Cas9 with single-nucleotide specificity and describe how it can be used to generate an in vivo mutation prevention system. Finally, I will outline a novel technology for the simultaneous generation of hundreds of mutant cells, each with a defined genetic alteration (point mutation, deletion or insertion), as well as methods for tracking the fitness of all generated mutants within a single experiment.
1. Current methods for Cas9-based transcriptional activation
2. Strategy to endow Cas9 with allele specificity
3. Describe our novel approach to generating and tracking the fitness of hundreds of mutant cell lines within a single experiment