Quantum dot‐mediated delivery of siRNA to inhibit sphingomyelinase activities in brain‐derived cells
Abstract
The use of RNAi to suppress protein synthesis offers a potential way of reducing the level of enzymes or the synthesis of mutant toxic proteins but there are few tools currently available for their delivery. To address this problem, bioconjugated quantum dots (QDs) containing a hydrophobic component (N‐palmitate) and a sequence VKIKK designed to traverse across cell membranes and visualize drug delivery were developed and tested on cell lines of brain origin. We used the Zn outer shell of the QD to bind HIS6 in JB577 (W•G•Dap(N‐Palmitoyl)•VKIKK•P9•G2•H6) and by a gel‐shift assay showed that siRNAs would bind to the positively charged KIKK sequence. By comparing many peptides and QD coatings, we showed that the QD‐JB577‐siRNA construct was taken up by cells of nervous system origin, distributed throughout the cytosol, and inhibited protein synthesis, implying that JB577 was also promoting endosome egress. By attaching siRNA for luciferase in a cell line over‐expressing luciferase, we showed 70% inhibition of mRNA after 24–48 h. To show more specific effects, we synthesized siRNA for neutral (NSMase2), acid (lysosomal ASMase) sphingomyelinase, and sphingosine kinase 1 (SK1), we demonstrated a dose‐dependent inhibition of activity. These data suggest that QDs are a useful siRNA delivery tool and QD‐siRNA could be a potential theranostic for a variety of diseases.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Oct 14, 2016
- Source ID
- 10.1111/jnc.13841
Entities
People
- Glyn Dawson
- Igor L. Medintz
- James B. Delehanty
- Jingdong Qin
- Kimihiro Susumu
- Philip E. Dawson
- Ted Getz
Organizations
- Defense Advanced Research Projects Agency
- Defense Threat Reduction Agency
- Office of Naval Research
- Scripps Research
- United States Naval Research Laboratory
- United States Public Health Service
- University of Chicago