Endosomal Escape and Delivery of CRISPR/Cas9 Genome Editing Machinery Enabled by Nanoscale Zeolitic Imidazolate Framework

J Am Chem Soc. 2018 Jan 10;140(1):143-146. doi: 10.1021/jacs.7b11754. Epub 2017 Dec 27.

Abstract

CRISPR/Cas9 is a combined protein (Cas9) and an engineered single guide RNA (sgRNA) genome editing platform that offers revolutionary solutions to genetic diseases. It has, however, a double delivery problem owning to the large protein size and the highly charged RNA component. In this work, we report the first example of CRISPR/Cas9 encapsulated by nanoscale zeolitic imidazole frameworks (ZIFs) with a loading efficiency of 17% and enhanced endosomal escape promoted by the protonated imidazole moieties. The gene editing potential of CRISPR/Cas9 encapsulated by ZIF-8 (CC-ZIFs) is further verified by knocking down the gene expression of green fluorescent protein by 37% over 4 days. The nanoscale CC-ZIFs are biocompatible and easily scaled-up offering excellent loading capacity and controlled codelivery of intact Cas9 protein and sgRNA.

MeSH terms

  • Animals
  • CHO Cells
  • CRISPR-Cas Systems / physiology*
  • Cricetulus
  • Endosomes / metabolism*
  • Gene Editing*
  • Imidazoles / chemistry*
  • Nanoparticles / chemistry*
  • Particle Size
  • Zeolites / chemistry*

Substances

  • Imidazoles
  • Zeolites