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Supplementary Materials

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  • fig. S1. Gel filtration of Cas9 complexes with AcrIIA4.
  • fig. S2. Exposed region analysis of SpyCas9 at AcrIIA4-free and AcrIIA4-bound states.
  • fig. S3. Cryo-EM of Cas9 ribonucleoprotein particles.
  • fig. S4. Classification and refinement workflow.
  • fig. S5. Atomic modeling.
  • fig. S6. Model comparison between AcrIIA4-bound and DNA-bound SpyCas9-sgRNA complexes.
  • fig. S7. Biological replicate data for BLI data shown in Fig. 3C.
  • fig. S8. EMSA of increasing concentrations of Cas9 binding to sgRNA in the absence or presence of AcrIIA4.
  • fig. S9. EMSA of Cas9-sgRNA binding to a target DNA with and without AcrIIA4.
  • fig. S10. Representative flow cytometry data used to create Fig. 4A.
  • fig. S11. Western blot of AcrIIA4-3XFLAG expression.
  • fig. S12. Representative flow cytometry data used to create the graph shown in Fig. 4B.
  • fig. S13. Representative flow cytometry data used to create the graph shown in Fig. 4C.
  • fig. S14. Quantification of on- and off-target editing at HBB, as measured by TIDE analysis.
  • table S1. Data collection and model refinement statistics.
  • table S2. Oligonucleotides used in this study.

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