Letter abstract
Nature Biotechnology 27, 567 - 571 (2009)
Published online: 17 May 2009 | doi:10.1038/nbt.1541
Efficient siRNA delivery into primary cells by a peptide transduction domain–dsRNA binding domain fusion protein
Akiko Eguchi1,2,3, Bryan R Meade1,2, Yung-Chi Chang4, Craig T Fredrickson2, Karl Willert2, Nitin Puri5 & Steven F Dowdy1,2
RNA interference (RNAi) induced by short interfering RNA (siRNA) allows for discovery research and large-scale screening1, 2, 3, 4, 5; however, owing to their size and anionic charge, siRNAs do not readily enter cells4, 5. 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.
- Howard Hughes Medical Institute, La Jolla, California, USA.
- Department of Cellular & Molecular Medicine, UCSD School of Medicine, La Jolla, California, USA.
- Japan Society for the Promotion of Science, Tokyo, Japan.
- Department of Pediatrics, UCSD School of Medicine, La Jolla, California, USA.
- Life Technologies, Austin, Texas, USA.
Correspondence to: Steven F Dowdy1,2 e-mail: sdowdy@ucsd.edu
MORE ARTICLES LIKE THIS
These links to content published by NPG are automatically generated.
RESEARCH
Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymeraseNature Letters to Editor (14 Apr 2005)
