CRISPR-guided DNA polymerase enabling diversification of all nucleotides in a tunable window
July 14-18, 2019
The capacity to diversify genetic codes advances our understanding and engineering of biological systems. A method to continuously diversify user-defined regions of a genome without requiring the integration of nucleic acid libraries would enable forward genetic approaches in systems not amenable to high efficiency homologydirected integration, rapid evolution of biotechnologically useful activity through accelerated and parallelized rounds of mutagenesis and selection, and cell lineage tracking. Here we developed EvolvR, the first system that can continuously diversify all nucleotides within a tunable window length at user-defined loci. Our results demonstrate that EvolvR enables multiplexed and continuous diversification of user-defined genomic loci that will be useful for a broad range of basic and biotechnological applications.
John Dueber, Shakked Halperin, David Schaffer, and Connor Tou, "CRISPR-guided DNA polymerase enabling diversification of all nucleotides in a tunable window" in "Biochemical and Molecular Engineering XXI", Christina Chan, Michigan State University, USA Mattheos Koffas, RPI, USA Steffen Schaffer, Evonik Industries, Germany Rashmi Kshirsagar, Biogen, USA Eds, ECI Symposium Series, (2019). https://dc.engconfintl.org/biochem_xxi/114