The CRISPR-Cas9 system is a powerful tool for gene editing. However, the low delivery efficiency of plasmids is still a huge obstacle to its widespread clinical application. Viral vectors can be used for efficient gene transfection, but it is easy to cause gene mutations; hydrodynamic injection or mechanical stimulation has a narrow application range for transfection; non-viral vectors (liposomes, DNA nanocoils, etc.) can be used to deliver Cas9 The complex of protein or mRNA and sgRNA, but the preparation of mRNA or protein requires tedious transcription and translation process, and is not stable enough, and the large size of plasmid makes delivery very difficult. Gold nanoparticles (AuNPs) have unique optical and structural properties and are good non-viral gene carriers. Their localized surface plasmon resonance (LSPR) effect can be used to trigger the release of drugs.
The Cas9-sgRNA expression plasmid targeting the Plk-1 gene that regulates mitosis was prepared, and then compressed into nuclear penetrating peptide modified gold nanoparticles through electrostatic interaction, and then wrapped in liposomes (DOTAP, DOPE, cholesterol). ), the outer layer is modified with PEG2000-DSPE to form a multifunctional carrier LACP. After the carrier enters the cancer tissue, the liposome guides it into the cancer cell, and the laser irradiation can make the gold nanoparticles produce a photothermal effect, and the temperature rises to release the penetrating peptide to guide the Cas9-sgRNA-Plk-1 plasmid into the nucleus, thereby completing the gene transfer Infection, induce gene editing, and make cancer cells apoptosis.
This multifunctional platform combines lipid carriers with gold nanoparticles for the thermal release of the CRISPR-Cas9 system to treat cancer. This method not only has certain clinical application potential, but also provides new ideas for gene transfection and editing in biological research.
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