Self-assembled DNA nanoclews for the efficient delivery of CRISPR-Cas9 for genome editing

Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12029-33. doi: 10.1002/anie.201506030. Epub 2015 Aug 27.


CRISPR-Cas9 represents a promising platform for genome editing, yet means for its safe and efficient delivery remain to be fully realized. A novel vehicle that simultaneously delivers the Cas9 protein and single guide RNA (sgRNA) is based on DNA nanoclews, yarn-like DNA nanoparticles that are synthesized by rolling circle amplification. The biologically inspired vehicles were efficiently loaded with Cas9/sgRNA complexes and delivered the complexes to the nuclei of human cells, thus enabling targeted gene disruption while maintaining cell viability. Editing was most efficient when the DNA nanoclew sequence and the sgRNA guide sequence were partially complementary, offering a design rule for enhancing delivery. Overall, this strategy provides a versatile method that could be adapted for delivering other DNA-binding proteins or functional nucleic acids.

Keywords: CRISPR-Cas9; DNA; drug delivery; genome editing; nanoparticles.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • CRISPR-Associated Proteins / administration & dosage*
  • CRISPR-Associated Proteins / chemistry
  • CRISPR-Associated Proteins / genetics
  • CRISPR-Cas Systems*
  • Cell Line
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • DNA / administration & dosage*
  • DNA / chemistry
  • DNA / genetics
  • Green Fluorescent Proteins / genetics
  • Humans
  • Mice, Nude
  • Nanoparticles / chemistry*
  • Neoplasms / genetics
  • Neoplasms / therapy
  • RNA, Guide / administration & dosage*
  • RNA, Guide / chemistry
  • RNA, Guide / genetics
  • Streptococcus pyogenes / chemistry
  • Streptococcus pyogenes / genetics


  • CRISPR-Associated Proteins
  • RNA, Guide
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • DNA