Efficient prime editing in two-cell mouse embryos using PEmbryo
Rebecca Kim, Ryan McNulty, Bradley Joyce, Antonio Mollica, Peter J. Chen, Purnima Ravisankar, Benjamin K. Law, David R. Liu, Jared E. Toettcher, Evgueni A. Ivakine, Eszter Pósfai, Britt Adamson
Abstract
Using transient inhibition of DNA mismatch repair during a permissive stage of development, we demonstrate highly efficient prime editing of mouse embryos with few unwanted, local byproducts (average 58% precise edit frequency, 0.5% on-target error frequency across 13 substitution edits at 8 sites), enabling same-generation phenotyping of founders. Whole-genome sequencing reveals that mismatch repair inhibition increases off-target indels at low-complexity regions in the genome without any obvious phenotype in mice.
Topics & Concepts
IndelBiologyEmbryoGeneticsComputational biologyGenomeGenome editingPhenotypePrime (order theory)DNACell biologyMolecular biologyGeneCombinatoricsGenotypeSingle-nucleotide polymorphismMathematicsCRISPR and Genetic EngineeringChromosomal and Genetic VariationsAdvanced biosensing and bioanalysis techniques