Efficient siRNA delivery into primary cells by a peptide transduction domain-dsRNA binding domain fusion protein.

Journal: 
Nat Biotechnol
Publication Year: 
2009
Authors: 
Akiko Eguchi , Bryan R Meade , Yung-Chi Chang , Craig T Fredrickson , Karl Willert , Nitin Puri , Steven F Dowdy
Public Summary: 
Scientific Abstract: 
RNA interference (RNAi) induced by short interfering RNA (siRNA) allows for discovery research and large-scale screening; however, owing to their size and anionic charge, siRNAs do not readily enter cells. Current approaches do not deliver siRNAs into a high percentage of primary cells without cytotoxicity. Here we report an efficient siRNA delivery approach that uses a peptide transduction domain-double-stranded RNA-binding domain (PTD-DRBD) fusion protein. DRBDs bind to siRNAs with high avidity, masking the siRNA's negative charge and allowing PTD-mediated cellular uptake. PTD-DRBD-delivered siRNA induced rapid RNAi in a large percentage of various primary and transformed cells, including T cells, human umbilical vein endothelial cells and human embryonic stem cells. We observed no cytotoxicity, minimal off-target transcriptional changes and no induction of innate immune responses. Thus, PTD-DRBD-mediated siRNA delivery allows efficient gene silencing in difficult-to-transfect primary cell types.

© 2013 California Institute for Regenerative Medicine