Active Intracellular Delivery of a Cas9/sgRNA Complex Using Ultrasound-Propelled Nanomotors.
Malthe Hansen-BruhnBerta Esteban-Fernández de ÁvilaMara Beltrán-GastélumJing ZhaoDoris E Ramírez-HerreraPavimol AngsantikulKurt Vesterager GothelfLiangfang ZhangJoseph WangPublished in: Angewandte Chemie (International ed. in English) (2018)
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.
Keyphrases
- crispr cas
- genome editing
- highly efficient
- induced apoptosis
- magnetic resonance imaging
- drug delivery
- cell cycle arrest
- cancer therapy
- computed tomography
- genome wide
- single cell
- mesenchymal stem cells
- endoplasmic reticulum stress
- cell therapy
- oxidative stress
- ultrasound guided
- hepatitis c virus
- bone marrow
- room temperature
- machine learning
- quantum dots
- transcription factor
- reduced graphene oxide
- men who have sex with men
- deep learning
- hiv testing