Active Intracellular Delivery of a Cas9/sgRNA Complex Using Ultrasound‐Propelled Nanomotors. Issue 10 (6th February 2018)

• Record Type: Journal Article
• Authors: Hansen‐Bruhn, Malthe; de Ávila, Berta Esteban‐Fernández; Beltrán‐Gastélum, Mara; Zhao, Jing; Ramírez‐Herrera, Doris E.; Angsantikul, Pavimol; Vesterager Gothelf, Kurt; Zhang, Liangfang; Wang, Joseph
• Is Part Of: Angewandte Chemie international edition. Volume 57:Issue 10(2018)
• Page Start: 2657
• Page End: 2661
• Publication Date: 2018-02-06
• Subjects: CRISPR/Cas9 -- GFP knockout -- intracellular delivery -- nanomotors -- nanowires; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201713082
• Languages: English
• ISSNs: 1433-7851

Abstract

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. Abstract : Speed of Sound : Ultrasound (US)‐powered motors enable the direct and rapid intracellular delivery of a fully functional Cas9/sgRNA complex. The acoustic nanomotors directly penetrate through the plasma membrane of GFP‐expressing B16F10 cells, releasing the carried Cas9/sgRNA inside the cell and achieving highly effective GFP gene knockout.