Pharmaceutical Research

, Volume 31, Issue 10, pp 2685–2695 | Cite as

Insight into Mechanisms of Cellular Uptake of Lipid Nanoparticles and Intracellular Release of Small RNAs

Research Paper
First Online:
Received:
Accepted:

Abstract

Purpose

Understanding mechanisms of cellular uptake and intracellular release would enable better design of nanocarriers for delivery of nucleic acids such as siRNA and microRNA (miRNA).

Method

In this study, we investigated cellular pharmacokinetics of siRNA by co-encapsulating fluorescently labeled siRNA and molecular beacon (MB) in four different formulations of cationic lipid nanoparticles (LNPs). A miRNA mimic was also used as a probe for investigating cellular pharmacokinetics, which correlated well with RNAi activities.

Results

We tried to find the best LNP formulation based on the combination of DOTMA and DODMA. When the DOTMA/DODMA ratio was at 5/40, the LNP containing a luciferase siRNA produced the highest gene silencing activity. The superior potency of DOTMA/DODMA could be attributed to higher uptake and improved ability to facilitate siRNA release from endosomes subsequent to uptake.

Conclusions

Our findings may provide new insights into RNAi transfection pathways and have implications on cationic LNP design.

KEY WORDS

intracellular trafficking lipid nanoparticles miRNA molecular beacon siRNA 
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Notes

ACKNOWLEDGMENTS AND DISCLOSURES

This work was supported by NSF Nanoscale Science and Engineering Center (NSEC) grant EEC-0914790 and the Chinese National 863 Project (No. 2012AA020804).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Bo Yu
    • 1
    • 2
  • Xinmei Wang
    • 2
  • Chenguang Zhou
    • 2
    • 3
  • Lesheng Teng
    • 3
    • 4
  • Wei Ren
    • 5
  • Zhaogang Yang
    • 3
  • Chih-Hsin Shih
    • 6
  • Tianyou Wang
    • 7
  • Robert J. Lee
    • 2
    • 3
    • 4
    • 10
  • Suoqin Tang
    • 8
  • L. James Lee
    • 1
    • 2
    • 9
  1. 1.Department of Chemical and Biomolecular EngineeringThe Ohio State UniversityColumbusUSA
  2. 2.NSF Nanoscale Science and Engineering Center (NSEC)The Ohio State UniversityColumbusUSA
  3. 3.Division of Pharmaceutics and Pharmaceutical Chemistry, College of PharmacyThe Ohio State UniversityColumbusUSA
  4. 4.College of Life SciencesJinlin UniversityChangchunPeople’s Republic China
  5. 5.Department of PhysiologyThe Ohio State UniversityColumbusUSA
  6. 6.Department of Chemical EngineeringFeng Chia UniversityTaichungTaiwan
  7. 7.Department of HematologyCapital Institute of Pediatrics Affiliated Children’s HospitalBeijingPeople’s Republic
  8. 8.Department of PediatricsPLA General HospitalBeijingPeople’s Republic China
  9. 9.The Ohio State UniversityColumbusUSA
  10. 10.ColumbusUSA

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