Fig. 4: In vivo lipid-mediated delivery of Cas9:sgRNA complexes restores hair bundle morphology.
a Experimental workflow. Samples were harvested, processed and imaged at 4 weeks after injection. b Images of scanning electron microscopy (SEM) of uninjected Atp2b2Obl/+ (left) and Cas9:Atp2b2-mut1 sgRNA: Lipo2000 injected Atp2b2Obl/+ (right) outer hair cell bundles. This was repeated independently for 3 times with similar results. Scale bar, 10 µm. c High-magnification images of outer hair cell bundles from the insets of (b) of uninjected Atp2b2Obl/+ (left) and Cas9:Atp2b2-mut1 sgRNA: Lipo2000 injected Atp2b2Obl/+ cochlea (right). The arrow points to shorter stereocilia in an outer hair cell from an injected ear that are missing from an outer hair cell from an uninjected ear. This was repeated independently for 3 times with similar results. Scale bar, 2 µm. d Images of scanning electron microscopy (SEM) uninjected Atp2b2Obl/+ (left) and Cas9:Atp2b2-mut1 sgRNA: Lipo2000 injected Atp2b2Obl/+ (right) inner hair cell bundles. This was repeated independently for 3 times with similar results. Scale bar, 10 µm. e High-magnification images of inner hair cell bundles from the insets of (d) of uninjected Atp2b2Obl/+ (left) and Cas9:Atp2b2-mut1 sgRNA: Lipo2000 injected Atp2b2Obl/+ cochlea (right). This was repeated independently for 3 times with similar results. Scale bar, 2 µm.
