Zsuzsa Bartos

2. Materials and Methods 2.1. Vector Constructs The p10-ABCG2-IRES2-GFP coding vector was used in all experiments that involved the expression of untagged ABCG2 variants. Mutations were introduced in the ABCG2 coding sequence by mutagenesis PCR and cassette change. All vector sequences were checked by Sanger sequencing. The p10-ABCG2-IRES2-GFP vector is a transposon coding vector.

Abstract Streptococcus pyogenes Cas9 (SpCas9) has been employed as a genome engineering tool with a promising potential within therapeutics. However, its off-target effects present major safety concerns for applications requiring high specificity. Approaches developed to date to mitigate this effect, including any of the increased-fidelity (i.e., high-fidelity) SpCas9 variants, only provide efficient editing on a relatively small fraction of targets without detectable off-targets.