Engineering Periodic shRNA for Enhanced Silencing Efficacy
Name
1-s2.0-S1525001616303744-main.pdf
Description
Published version
Size
1.34 MB
Format
Adobe PDF
Checksum (MD5)
65b91138bcdcea21d795cb56e5428a28
Author(s) • •
Wu, Connie
Shopsowitz, Kevin E
Hammond, Paula T
Date Issued
2016
Journal
Molecular Therapy
Publisher
Elsevier BV
Version
Final published version
Abstract
RNA interference (RNAi) provides a versatile therapeutic approach via silencing of specific genes, particularly undruggable targets in cancer and other diseases. However, challenges in the delivery of small interfering RNA (siRNA) have hampered clinical translation. Polymeric or periodic short hairpin RNAs (p-shRNAs) - synthesized by enzymatic amplification of circular DNA - are a recent development that can potentially address these delivery barriers by showing improved stability and complexation to enable nanoparticle packaging. Here, we modify these biomacromolecules via structural and sequence engineering coupled with selective enzymatic digestion to generate an open-ended p-shRNA (op-shRNA) that is cleaved over ten times more efficiently to yield siRNA. The op-shRNA induces considerably greater gene silencing than p-shRNA in multiple cancer cell lines up to 9 days. Furthermore, its high valency and flexibility dramatically improve complexation with a low molecular weight polycation compared to monomeric siRNA. Thus, op-shRNA provides an RNAi platform that can potentially be packaged and efficiently delivered to disease sites with higher therapeutic efficacy.
MIT Department
Koch Institute for Integrative Cancer Research at MIT
Massachusetts Institute of Technology. Department of Chemical Engineering
Massachusetts Institute of Technology. Institute for Soldier Nanotechnologies
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1038/MT.2016.69