Products/Services Used | Details | Operation |
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Gene Synthesis> | … 6,15 Constructs used in this study were generated by Gibson assembly, or synthesized by Genscript. Gene fragments used for polymerase chain reaction (PCR) were purchased as mammalian codon-optimized gene fragments from Genscript … | Get A Quote |
Base editors are fusions of a deaminase and CRISPR-Cas ribonucleoprotein that allow programmable installment of transition mutations without double-strand DNA break intermediates. The breadth of potential base editing targets is frequently limited by the requirement of a suitably positioned Cas9 protospacer adjacent motif. To address this, we used structures of Cas9 and TadA to design a set of inlaid base editors (IBEs), in which deaminase domains are internal to Cas9. Several of these IBEs exhibit shifted editing windows and greater editing efficiency, enabling editing of targets outside the canonical editing window with reduced DNA and RNA off-target editing frequency. Finally, we show that IBEs enable conver... More