Active Intracellular Delivery of a Cas9/sgRNA Complex Using Ultrasound-Propelled Nanomotors

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  • Malthe Hansen-Bruhn
  • ,
  • Berta Esteban Fernández de Ávila, University of California, San Diego
  • ,
  • Mara Beltrán-Gastélum, University of California, San Diego
  • ,
  • Jing Zhao, University of California, San Diego
  • ,
  • Doris E. Ramírez-Herrera, University of California, San Diego
  • ,
  • Pavimol Angsantikul, University of California, San Diego
  • ,
  • Kurt Vesterager Gothelf
  • Liangfang Zhang, University of California, San Diego
  • ,
  • Joseph Wang, University of California, San Diego

Direct and rapid intracellular delivery of a functional Cas9/sgRNA complex using ultrasound-powered nanomotors is reported. The Cas9/sgRNA complex is loaded onto the nanomotor surface through a reversible disulfide linkage. A 5 min ultrasound treatment enables the Cas9/sgRNA-loaded nanomotors to directly penetrate through the plasma membrane of GFP-expressing B16F10 cells. The Cas9/sgRNA is released inside the cells to achieve highly effective GFP gene knockout. The acoustic Cas9/sgRNA-loaded nanomotors display more than 80 % GFP knockout within 2 h of cell incubation compared to 30 % knockout using static nanowires. More impressively, the nanomotors enable highly efficient knockout with just 0.6 nm of the Cas9/sgRNA complex. This nanomotor-based intracellular delivery method thus offers an attractive route to overcome physiological barriers for intracellular delivery of functional proteins and RNAs, thus indicating considerable promise for highly efficient therapeutic applications.

Original languageEnglish
JournalAngewandte Chemie - International Edition
Pages (from-to)2657-2661
Number of pages5
Publication statusPublished - 1 Mar 2018

    Research areas

  • CRISPR/Cas9, GFP knockout, intracellular delivery, nanomotors, nanowires

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