Figure 1. AcrIIA2 Selectively Forms a Stable Complex with the sgRNA-Loaded S. pyogenes Cas9.
(A) Flow chart for reconstitution and isolation of the AcrIIA2-bound SpyCas9-sgRNA ternary complex using ultrafiltration combined with size-exclusion chromatography (UF-SEC).
(B) Size-exclusion chromatogram ofSpyCas9-AcrIIA2 in the presence or absence of sgRNA. The inset representsthe calibration curve obtained using standard molecular weight markers.
(C) Coomassie-stained polyacrylamide gel showing the co-purification of AcrIIA2 with sgRNA-bound SpyCas9.