Science Advances

Supplementary Materials

The PDFset includes:

  • Fig. S1. Coexpressing mild doses of Acr with Cas9 improves genome editing specificity.
  • Fig. S2. Kinetic insulation of ON- and OFF-target editing events upon AAV-mediated delivery.
  • Fig. S3. Kinetic insulation of ON- and OFF-target editing upon transient transfection is difficult for the HBB sgRNA.
  • Fig. S4. Screen of AcrIIA4 variants weakened by domain insertion.
  • Fig. S5. Screen of Cas-Acr fusion variants.
  • Fig. S6. Evaluation of editing specificity with Cas-Acr fusions.
  • Fig. S7. Targeted amplicon sequencing analysis of Cas-Acr editing specificity.
  • Fig. S8. Comparison of Cas-Acr fusions to other Cas9 high-fidelity variants by TIDE sequencing.
  • Fig. S9. Kinetics of Cas9 and Acr expression and corresponding ON- and OFF-target editing.
  • Fig. S10. Estimation of Cas9, AcrIIA4, and Cas-Acr protein levels and model fits.
  • Fig. S11. Kinetic insulation of ON- and OFF-target editing events by Cas-Acr variants.
  • Fig. S12. Supplementary gel images.
  • Table S1. Equations of the Cas9 gene editing model visualized in Fig. 1A.
  • Table S2. Overview of AcrIIA4-LOV2 hybrids from Bubeck et al. ( 29) used in this study.
  • Table S3. Plasmids used and constructed in this study.
  • Table S4. sgRNA ON- and OFF-target sequences.
  • Table S5. Primers used for genomic PCRs.
  • Table S6. Equations of the Cas-Acr gene editing model visualized in Fig. 4A.
  • Table S7. Parameter estimates and confidence intervals.

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Other Supplementary Material for this manuscript includes the following:

  • Data file S1 (.zip format). Annotated vector sequences (GenBank files).
  • Data file S2 (.zip format). Scripts for model simulations in MATLAB.

Files in this Data Supplement: