Efficient single and combinatorial CRISPR gene editing with 3Cs gRNAs.

Abstract

Current methods to generate single or combinatorial gene perturbation libraries are labor intense and require multiple ligation and cloning steps. Moreover, increasing sequence diversity negatively affects sequence distribution, resulting in biased libraries and large cell culture demands. I will present our recently developed 3Cs technology for the rapid and cloning-free generation of single and combinatorial gRNA libraries. Due to their low sequence bias, these reagents can be applied with minimal experimental coverages and reduce cell culture demands by at least 10-fold. Despite carefully explaining the 3Cs technology, I will also provide two real-world examples of how my lab applies the 3Cs technology to explore single and combinatorial gene effects.

Biography

Manuel studied Biotechnology and obtained his PhD from the Max Planck Institute of Biochemistry and the Biozentrum in Basel. For his postdoc, he joined the group of Steven Dowdy at the University of California in San Diego, where he became interested in gene editing technologies. Since December 2015, Manuel is a group leader at the Institute of Biochemistry II at the Medical Faculty of the Goethe University Frankfurt, Germany. His laboratory develops gene editing technologies and applies them to understand the various aspects of cellular transformation and drug resistance. A major achievement of his lab is the development of the 3Cs technology that recently culminated in the foundation of the university spin-off Vivlion for which Manuel is the Chief Scientific Officer.