Article
CasPER, a method for directed evolution in genomic contexts using mutagenesis and CRISPR/Cas9.
Metabolic engineering - 1 Jul 2018
Jakočiūnas Tadas, Pedersen Lasse E, Lis Alicia V, Jensen Michael K, Keasling Jay D
Abstract excerpt
Here we describe a method for robust directed evolution using mutagenesis of large sequence spaces in their genomic contexts. The method employs error-prone PCR and Cas9-mediated genome integration of mutant libraries of large-sized donor variants into single or multiple genomic sites with efficiencies reaching 98-99%. From sequencing of genome integrants, we determined that the mutation frequency along the donor...
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