Fig. 5.

APC-mediated optogenetic control of programmable genome editing in vivo. (A) Schematic illustration (Left). Transfection with APC–HSP-Cas9 complexes, and then subcutaneous implantation. Whereas the right implanted position was exposed to irradiation from 5 to 30 min, the left position (nonirradiation) was used as the control. In vivo luciferase expression (Right). (B) Indel mutations detected by T7E1 assay. (C) Quantitative analysis of indel mutations. Mean ± SD; n = 3. (D) Schematic illustration (Left). APC–HSP-Cas9 complexes were subcutaneously injected into the muscle of the hind limb of BALB/c mice, followed by irradiation at 1,064 nm. In vivo luciferase expression (Right). (E) Indel mutations detected by T7E1 assay. (F) Quantitative analysis of indel mutations. Mean ± SD; n = 3. (G) Schematic illustration (Left). Tumor-bearing mice were administered APC–HSP-Cas9 complexes through peritumoral injection, and the tumor (Right) was then exposed to irradiation for 30 min. In vivo luciferase expression (Middle). (H) Indel mutations detected by T7E1 assay. (I) Quantitative analysis of indel mutations. Mean ± SD; n = 3. (J) Evaluation of in vivo luciferase expression through intravenous injection, followed by optogenetic activation in the liver. Schematic illustration (Left). BALB/c mice were injected with APC–HSP-Cas9 complexes via tail-vein injection, and the liver position was exposed to irradiation for 30 min after the transfection. (K) Indel mutations from different tissues after the systemic administration of nanoCRISPR through intravenous injection, followed by injection with APC–HSP-Cas9 complexes via tail-vein injection, and the liver position was exposed to the irradiation for 30 min after the transfection. (L) Optogenetic regulation of transcriptional activation of mCherry expression through in vivo transfection and optogenetic activation. The arrowheads in B, E, H, and K show the cleaved DNA fragments of the target genome.