Journal: Cell Reports Medicine
Article Title: Specific macrophage RhoA targeting CRISPR-Cas9 for mitigating osteoclastogenesis-induced joint damage in inflammatory arthritis
doi: 10.1016/j.xcrm.2025.102046
Figure Lengend Snippet: Targeting effect of SMART-Cas9 in vivo by TP fluorescence imaging (A) Schematic illustration of CIA wild-type mouse paw joints imaged by TP excitation fluorescence microscopy after injection with SMART-Cas9-CM-Dil. (B) Representative TP time-lapse images of paw joints in CIA mice after intravenous injection with SMART-Cas9-CM-Dil. Scale bar, 10 μm. (C) Movement tracks of immunocytes after engulfing SMART-Cas9-CM-Dil in CIA mouse paw joints tissue. The green and red indicate collagen and CM-Dil, respectively. Scale bar, 10 μm. (D) DNA electrophoresis identification of CX3CR-1 GFP/+ C57BL/6 mice. (E) Schematic illustration of CX3CR-1 GFP/+ CIA mice joints imaged by TP excitation fluorescence microscopy. (F) 3D TP images of paw joints in CX3CR-1 GFP/+ healthy mice and saline- and SMART-Cas9-CM-Dil-injected CX3CR-1 GFP/+ CIA mice. Scale bar, 100 μm. (G) TP images of paw joints of CX3CR-1 GFP/+ CIA mice after intravenous injection with saline or SMART-Cas9-CM-Dil. Scale bar, 200 μm. (H) Spearman’s correlation between GFP and CM-Dil. (I) Time-lapse TP images of paw joints of CX3CR-1 GFP/+ CIA mice after intravenous injection with SMART-Cas9-CM-Dil. Scale bar, 200 μm. Red arrows indicate the macrophages that engulf the SMART-Cas9-CM-Dil; the white circles indicate new emerging macrophages after intravenous injection with SMART-Cas9-CM-Dil.
Article Snippet: A commercial cell membrane dye, CM-Dil, was employed to label the SMART-Cas9, and imaging was conducted using two-photon excitation fluorescence microscopy (A1R-MP, Nikon) in vivo .
Techniques: In Vivo, Fluorescence, Imaging, Microscopy, Injection, Nucleic Acid Electrophoresis, Saline