Thermo-triggered Release of CRISPR-Cas9 System by Lipid-Encapsulated Gold Nanoparticles for Tumor Therapy

Angew Chem Int Ed Engl. 2018 Feb 5;57(6):1491-1496. doi: 10.1002/anie.201708689. Epub 2018 Jan 15.

Abstract

CRISPR/Cas9 system is a powerful toolbox for gene editing. However, the low delivery efficiency is still a big hurdle impeding its applications. Herein, we report a strategy to deliver Cas9-sgPlk-1 plasmids (CP) by a multifunctional vehicle for tumor therapy. We condensed CPs on TAT peptide-modified Au nanoparticles (AuNPs/CP, ACP) via electrostatic interactions, and coated lipids (DOTAP, DOPE, cholesterol, PEG2000-DSPE) on the ACP to form lipid-encapsulated, AuNPs-condensed CP (LACP). LACP can enter tumor cells and release CP into the cytosol by laser-triggered thermo-effects of the AuNPs; the CP can enter nuclei by TAT guidance, enabling effective knock-outs of target gene (Plk-1) of tumor (melanoma) and inhibition of the tumor both in vitro and in vivo. This AuNPs-condensed, lipid-encapsulated, and laser-controlled delivery system provides a versatile method for high efficiency CRISPR/Cas9 delivery and targeted gene editing for treatment of a wide spectrum of diseases.

Keywords: CRISPR/Cas9 delivery; gold nanoparticles; lipids; photothermal effect; tumor therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / radiation effects
  • CRISPR-Associated Protein 9 / genetics*
  • Cell Cycle Proteins / genetics
  • Cell Line, Tumor
  • Gene Transfer Techniques
  • Glutathione / chemistry
  • Gold / chemistry*
  • Humans
  • Hyperthermia, Induced
  • Lasers
  • Lipids / chemistry*
  • Melanoma, Experimental / pathology
  • Melanoma, Experimental / therapy*
  • Metal Nanoparticles / chemistry*
  • Mice
  • Microscopy, Confocal
  • Peptide Fragments / chemistry
  • Plasmids / genetics
  • Plasmids / therapeutic use*
  • Polo-Like Kinase 1
  • Protein Serine-Threonine Kinases / genetics
  • Proto-Oncogene Proteins / genetics
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • Surface Plasmon Resonance

Substances

  • Cell Cycle Proteins
  • Lipids
  • Peptide Fragments
  • Proto-Oncogene Proteins
  • RNA, Guide, CRISPR-Cas Systems
  • Gold
  • Protein Serine-Threonine Kinases
  • CRISPR-Associated Protein 9
  • Glutathione