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. Author manuscript; available in PMC: 2023 Aug 4.
Published in final edited form as: Mol Cell. 2022 Jul 13;82(15):2754–2768.e5. doi: 10.1016/j.molcel.2022.06.007

Figure 1. Pfu Cascade-Cas3 form an integral effector complex.

Figure 1.

(A) Arrangement of the Pyrococcus furiosus Type I-A CRISPR-cas operon. (B) Agarose EMSA showing that Pfu Cas3 significantly improved the DNA target binding behavior of Pfu Cascade. (C) Agarose EMSA using differentially labeled binding partners to reveal that I-A Pfu Cas3 bound constitutively to Pfu Cascade. In contrast, I-E Tfu Cas3 only interacted with Tfu Cascade upon full R-loop formation. Cas3, Cascade, and dsDNA were labeled with Cy3 (rendered in green), Cy5 (red), and FAM (blue), respectively. Overlapping colors indicate complex formation that survived electrophoresis.