Format

Send to

Choose Destination
Angew Chem Int Ed Engl. 2018 Mar 1;57(10):2657-2661. doi: 10.1002/anie.201713082. Epub 2018 Feb 6.

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

Author information

1
University of California San Diego, 9500 Gilman Drive, La Jolla, CA, 92093, USA.
2
Interdisciplinary Nanoscience Center (iNANO) and Department of Chemistry, Aarhus University, 8000, Aarhus C, Denmark.

Abstract

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.

KEYWORDS:

CRISPR/Cas9; GFP knockout; intracellular delivery; nanomotors; nanowires

PMID:
29325201
DOI:
10.1002/anie.201713082

Supplemental Content

Full text links

Icon for Wiley
Loading ...
Support Center