Skip to main content
. Author manuscript; available in PMC: 2023 Mar 27.
Published in final edited form as: Science. 2022 Aug 25;377(6612):1278–1285. doi: 10.1126/science.add5064

Fig. 3. Target validation and cleavage mechanisms by Sb-gRAMP RNP.

Fig. 3.

(A) Models depicting the gate closed structure in resting state (left) and gate open structure in target RNA bound state (middle). Superposition is shown as the right panel. (B) Sequence alignment at the gating loop region. Conserved residues are highlighted in burgundy red. (C) Structural comparison of the resting and non-matching PFS RNA bound states. Vector length is proportional to residue movement distance. Hinge motion in Cas11 is pronounced. (D) EMSA (top) and urea-PAGE (bottom) to evaluate the impact of gating loop disruption on the binding and cleavage of partially matching RNA targets. (E) Mechanistic model depicting the essential role of the gating loop in target validation. (F) Structural basis for Site 1 cleavage. (G) Structural basis for Site 2 cleavage. (H) Impact of Site 1 (in blue) and Site 2 (in black) mutations on target RNA cleavage efficiency.