Martin Jinek

CRISPR-Cas genome editors: structures, mechanisms and applications

Abstract

CRISPR-associated effector nucleases have been repurposed as powerful tools for precision genome editing in eukaryotic cells and organisms. The CRISPR-associated protein Cas9 functions as an RNA-guided, programmable DNA-cutting enzyme. My research focuses on studying the molecular mechanisms of Cas9 and other CRISPR-associated nucleases using a combination of structural, biochemical and biophysical approaches. To this end, we initially determined the three-dimensional structures of Cas9 in complex with a guide RNA and target DNA, revealing atomic interactions underpinning the recognition of the protospacer adjacent motif (PAM), which is necessary to facilitate strand separation in the DNA and guide RNA hybridization. We subsequently focused on Cas12a (Cpf1), an alternative genome editor nuclease. Structures of guide- and target-bound Cas12a complexes and corroborating biochemical studies revealed that Cas12a preorganizes the seed sequence of the guide RNA to facilitate target DNA recognition and uses a sequential mechanism to cleave both strands of the target DNA using a single nuclease active site. Our most recent studies aim to understand the molecular basis for the off-target activity of Cas9, thereby contributing to the development of off-target prediction algorithms and engineered high-fidelity Cas9 variants. Collectively, these studies provide the mechanistic foundation for understanding the molecular function of CRISPR-based genome editor nucleases and for the on-going development of CRISPR-Cas genetic engineering for biotechnological and therapeutic applications.

 


Biography

Martin Jinek was born in  Czechoslovakia in 1979. He studied Chemistry and Natural Sciences at Trinity College, University of Cambridge (UK). In 2006, he received his PhD from the European Molecular Biology Laboratory (EMBL) in Heidelberg where he conducted his thesis in the lab of Dr. Elena Conti. He then moved to the University of California in Berkeley for postdoctoral research with Prof. Jennifer Doudna. Martin joined the Department of Biochemistry in February 2013 as Assistant Professor on tenure track.

He is interested in the molecular mechanisms that orchestrate cellular regulation through protein-RNA interactions. His studies include biochemical and structural approaches to investigate these processes at the atomic level.

Bio

University of Zurich

Professor at the department of Biochemistry