Repurposing CRISPR for versatile mammalian genome engineering and imaging
Dr. Lei (Stanley) Qi
Center for Systems Biology, UCSF
February 20, 2014
Foege N130A, Wallace H. Coulter Seminar Room
The functional interrogation and genetic reprogramming of cell fate and identity require efficient genome engineering technologies. The bacterial immune system CRISPR has emerged as a powerful approach for directed genome editing in diverse organisms via an RNA-guided Cas9 nuclease. In addition to its utility in genome editing, we have repurposed the CRISPR system as a general platform with several versatile applications, including transcriptional regulation, epigenetic modification, and chromatin imaging. We pioneered the use of a catalytically inactive version of Cas9 (dCas9), which lacks nuclease activity, and coupled dCas9 to transcription factors or chromatin modifiers for tunable and multiplexed endogenous gene expression or repression. Furthermore, fusing dCas9 to EGFP allows the site-specific visualization of chromatin structure and dynamics in living human cells. Our work establishes a set of powerful and versatile CRISPR RNA-guided technologies for efficient, robust, and multiplexable human genome engineering and imaging.
Dr. Qi received his Ph.D. in Bioengineering from UC Berkeley, where he studied synthetic and systems biology with Dr. Adam Arkin and Dr. Jennifer Doudna. As a graduate student, he focused on understanding the design principles of engineering and applying synthetic noncoding RNA regulators to build transcriptional and translational genetic circuits. Dr. Qi started his lab at UCSF as an independent fellow in 2012. His lab has developed CRISPR genome engineering technologies for precise gene regulation and chromatin imaging. The new technologies likely provide superior alternatives to existing methods base on TALENS or RNA interference for genome and cell engineering. He is currently developing customizable therapeutically strategies through non-invasive genetic or epigenetic modifications to reliably detect and correct disease states.