Abstract
Gene expression regulation by small interfering RNA (siRNA) holds promise in treating a wide range of diseases through selective gene silencing. However, successful clinical application of nucleic acid-based therapy requires novel delivery options. Herein, to achieve efficient delivery of negatively charged siRNA duplexes, the internal cavity of “humanized” chimeric Archaeal ferritin (HumAfFt) was specifically decorated with novel cationic piperazine-based compounds (PAs). By coupling these rigid-rod-like amines with thiol-reactive reagents, chemoselective conjugation was efficiently afforded on topologically selected cysteine residues properly located inside HumAfFt. The capability of PAs-HumAfFt to host and deliver siRNA molecules through human transferrin receptor (TfR1), overexpressed in many cancer cells, was explored. These systems allowed siRNA delivery into HeLa, HepG2, and MCF-7 cancer cells with improved silencing effect on glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expression with respect to traditional transfection methodologies and provided a promising TfR1-targeting system for multifunctional siRNA delivery to therapeutic applications.
Highlights
Small interfering RNA represents a revolutionary tool for gene therapy with a wide array of potential applications in the regulation of gene expression
We further explored the possibility to employ PAsHumAfFt as as safe and effective targeting shuttle of noncovalently loading Small interfering RNA (siRNA) for mediating the downregulation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expression, a housekeeping gene implicated in the catalysis of an important energy step in carbohydrate metabolism
As predicted by theoretical pKa values calculated through an ab initio quantum chemical program (Jaguar), the amine groups may significantly contribute to the electrostatic attraction and incorporation of siRNA molecules into the HumAfFt at a physiological pH (Figure S26)
Summary
Small interfering RNA (siRNA) represents a revolutionary tool for gene therapy with a wide array of potential applications in the regulation of gene expression. Protein-mediated siRNA delivery has several advantages such as facile chemical modifications and good biocompatibility, which may overcome various hurdles associated with efficient siRNA delivery.[7,8] Ferritin nanocages emerged as ideal delivery systems endowed with a well-defined hollow spherical architecture with inner and outer diameters of 8 and 12 nm, respectively, and precisely self-assembled from 24 copies of identical 20 kDa subunits These systems can be and accurately manipulated by genetic modifications to enhance their loading cargo properties with appropriate chemical bioconjugations.[9−13] Ferritin nanocages display homogeneity, low production costs, improved thermal stability, and cellular uptake activity of small bioactive compounds.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
