Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA

RNA-guided endonucleases (RGENs) derived from the CRISPR/Cas system represent an efficient tool for genome editing. RGENs consist of two components: Cas9 protein and guide RNA. Plasmid-mediated delivery of these components into cells can result in uncontrolled integration of the plasmid sequence into the host genome, and unwanted immune responses and potential safety problems that can be caused by the bacterial sequences. Furthermore, this delivery method requires transfection tools. Here we show that simple treatment with cell-penetrating peptide (CPP)-conjugated recombinant Cas9 protein and CPP-complexed guide RNAs leads to endogenous gene disruptions in human cell lines. The Cas9 protein was conjugated to…

• NR
  National Research Foundation
• HU
  Hanyang University
• YU
  Yonsei University
• KB
  Korea Basic Science Institute
• NR
  National Research Foundation of Korea

  Awards: 2013M3A9B4076544, 2011-0019357
• KA
  Korea Advanced Institute of Science and Technology