FIGURE 9.

NIR-II/CRISPR-Cas9-based imaging systems. (A) Diagram of SPPF-Dex mediated CRISPR-Cas9 imaging, and illustration of the intracellular genome editing process upon 808 nm laser irradiation. Upon the NIR-II laser irradiation, the distribution of SPPF-Dex can be tracked in vivo. Simultaneously, the photothermal conversion of SPPF facilitates the endolysosomal escape of SPPF-Dex and release of CRISPR-Cas9. Dex dilates the nuclear pores to initiate the translocation of CRISPR-Cas9 for genome editing. (B) Agarose gel retardation assay of SPPF/pDNAs, morphologies of SPPF, SPPF/pDNA, and SPPF/pDNA after 808 nm laser irradiation detected by TEM showed high efficiency as a carrier. (C) GFP gene disruption efficacy after different treatment in engineered GFP labeled cells detected by confocal microscope and flow cytometry. (D) In vivo NIR-II imaging of the injection of SPPF-Dex/Cas9-GFP and in vitro NIR-II imaging of the tumor and other organs harvested to analysis of the in vivo distribution. (E) T7E1 assay was carried out to indicate indel mutations of PCR products of GFP gene retrieved from GFP labeled tumor with different treatment. Histological examination of H&E staining of spleen and liver sections showed a minimal side effect. G1, GFP labeled tumor treated with PBS and 808 nm laser irradiation. G2, GFP labeled tumor treated with SPPF-Dex/Cas9-sgGFP. G3, GFP labeled tumor treated with SPPF-Dex/Cas9-sgGFP and 808 nm laser irradiation (reproduced from (Li et al., 2019) with permission from Advanced materials (Deerfield Beach, Fla.)).