Efficacy of chitosan/double-stranded RNA polyplex nanoparticles for gene silencing under variable environmental conditions

Stuart S. Lichtenberg, Kanthi Nuti, Jason DeRouchey, Olga V. Tsyusko, Jason M. Unrine

Research output: Contribution to journalArticlepeer-review

9 Scopus citations


We investigated the ability of chitosan/double-stranded RNA polyplex nanoparticles to silence genes inCaenorhabditis elegansin different environmentally analogous media. Using fluorescence microscopy, we were able to rapidly assess gene knockdown and dsRNA uptake under numerous conditions. Scanning transmission electron micrographs of polyplexes confirms heterogeneous distribution of chitosan and RNA in single particles and a wide range of particle morphologies. High pH and the presence of natural organic matter inhibited the ability of polyplex nanoparticles to silence genes, but were unaffected by the presence of inorganic nitrate and phosphate. Environmental media did not affect particle size in any specific pattern, as determined by dynamic light scattering and fluorescence correlation spectroscopy. The efficacy of polyplexes seems to be closely tied to zeta potential, as all treatments that resulted in a net negative zeta potential (high pH and high natural organic matter) failed to achieve gene knockdown. These results support earlier work that emphasized the importance of charge in gene carriers and will aid in the development of effective gene silencing biological control agents.

Original languageEnglish
Pages (from-to)1582-1592
Number of pages11
JournalEnvironmental Science: Nano
Issue number5
StatePublished - May 2020

Bibliographical note

Funding Information:
The authors wish to thank S. Shrestha and A. Wamucho for assistance with C. elegans. All strains were obtained from the Caenorhabditis Genetics Center, which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). This material is based upon work supported by the National Science Foundation under Grant No. CBET-1712323 and Cooperative Agreement EF-0830093. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the author(s) and do not necessarily reflect the views of the National Science Foundation. Portions of the work were also supported by the U.S.-Israel Agricultural Research and Development Fund through grant number IS-4964-16 R.

Publisher Copyright:
© The Royal Society of Chemistry 2020.

ASJC Scopus subject areas

  • Materials Science (miscellaneous)
  • Environmental Science (all)


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