Products/Services Used | Details | Operation |
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GenParts™ DNA Fragments> | … The “PE3” system differs from PE2 by using an additional single guide RNA (sgRNA) to nick the non-edited strand at a location away from … were designed using GenSmart Codon Optimization (Genscript) and ordered as gBlock gene fragments (Integrated DNA Technologies). … | Get A Quote |
While prime editing enables precise sequence changes in DNA, cellular determinants of prime editing remain poorly understood. Using pooled CRISPRi screens, we discovered that DNA mismatch repair (MMR) impedes prime editing and promotes undesired indel byproducts. We developed PE4 and PE5 prime editing systems in which transient expression of an engineered MMR-inhibiting protein enhances the efficiency of substitution, small insertion, and small deletion prime edits by an average 7.7-fold and 2.0-fold compared to PE2 and PE3 systems, respectively, while improving edit/indel ratios by 3.4-fold in MMR-proficient cell types. Strategic installation of silent mutations near the intended edit can enhance prime editing... More