Efficient Delivery of Genome-Editing Proteins In Vitro and In Vivo

Details

• Alternative Title:
  Nat Biotechnol
• Personal Author:
  Zuris, John A. ; Thompson, David B. ; Shu, Yilai ; Guilinger, John P. ; Bessen, Jeffrey L. ; Hu, Johnny H. ; Maeder, Morgan L. ; Joung, J. Keith ; Chen, Zheng-Yi ; Liu, David R.
• Description:
  Efficient intracellular delivery of proteins is needed to fully realize the potential of protein therapeutics. Current methods of protein delivery commonly suffer from low tolerance for serum, poor endosomal escape and limited in vivo efficacy. Here we report that common cationic lipid nucleic acid transfection reagents can potently deliver proteins that are fused to negatively supercharged proteins, that contain natural anionic domains or that natively bind to anionic nucleic acids. This approach mediates the potent delivery of nM concentrations of Cre recombinase, TALE- and Cas9-based transcription activators, and Cas9:sgRNA nuclease complexes into cultured human cells in media containing 10% serum. Delivery of unmodified Cas9:sgRNA complexes resulted in up to 80% genome modification with substantially higher specificity compared to DNA transfection. This approach also mediated efficient delivery of Cre recombinase and Cas9:sgRNA complexes into the mouse inner ear in vivo, achieving 90% Cre-mediated recombination and 20% Cas9-mediated genome modification in hair cells.
• Subjects:
  Article Cations In Vitro Techniques Lipids Proteins Trans-Activators Transfection
• Source:
  Nat Biotechnol. 33(1):73-80.
• Pubmed ID:
  25357182
• Pubmed Central ID:
  PMC4289409
• Document Type:
  Journal Article
• Funding:
  DP1 GM105378/DP/NCCDPHP CDC HHS/United States ; F32 GM 106601-2/GM/NIGMS NIH HHS/United States ; R01 DC006908/DC/NIDCD NIH HHS/United States ; R01 DC006908/DC/NIDCD NIH HHS/United States ; R01 GM095501/GM/NIGMS NIH HHS/United States ; R01 GM095501/GM/NIGMS NIH HHS/United States ; Howard Hughes Medical Institute/United States ; Howard Hughes Medical Institute/United States
• Volume:
  33
• Issue:
  1
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:9d2535d61b7b97dd825d8703d07b032b9b5ae8cf9e873c532bc64c8ffcf9cec8
• Download URL:
  https://stacks.cdc.gov/view/cdc/30227/cdc_30227_DS1.pdf
• File Type:
  [PDF - 1010.36 KB ]