Oxime Ether Lipids as Transfection Agents: Assembly and Complexation with siRNA

  • Anu Puri
  • Serena Zampino
  • Mathias Viard
  • Bruce A. Shapiro
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1632)

Abstract

RNAi-based therapeutic approaches to combat cancer and other diseases are currently an area of great interest. However, practical applications of this approach rely on optimal tools to carry and deliver siRNA to the desired site. Oxime ether lipids (OELs) are a class of molecules among other various carriers being examined for siRNA delivery. OELs, relatively new candidates, belong to a class of non-glycerol based lipids and have begun to claim their place as an siRNA delivery carrier in the field of RNAi therapy. Chemical synthesis steps of OELs are considered relatively simple with the ability to modify the functionalities as desired. OEL–siRNA complexes can be assembled in the presence of serum-containing buffers (or cell culture media) and recent data from our and other groups have demonstrated that OELs are viable carriers for siRNA delivery in the cell culture systems. In this chapter, we provide the details of experimental protocols routinely used in our laboratory to examine OEL–siRNA complexes including their assembly, stability, and transfection efficiencies.

Key words

Oxime ether lipids RNAi therapy Nucleic acid delivery oxime ether lipids Structure–activity relationship Nonsymmetric hydrophobic domain Lipoplexes RNA interference Breast cancer cells 

Notes

Acknowledgments

This research has been funded in whole or in part with Federal funds from the Frederick National Laboratory for Cancer Research, National Institutes of Health, under contract HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does the mention of trade names, commercial products, or organizations imply endorsement by the US Government. This work was supported by the Intramural Research Program of the National Institutes of Health, Center for Cancer Research, the National Cancer Institute. We thank Dr. Kshitij Gupta (Indian Institute of Science, Bangalore) and Dr. Michael Nantz (the University of Louisville, Kentucky) for their critical role in the exploration of OELS for siRNA delivery.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Anu Puri
    • 1
  • Serena Zampino
    • 1
  • Mathias Viard
    • 1
    • 2
  • Bruce A. Shapiro
    • 1
  1. 1.RNA Structure and Design Section, RNA Biology Laboratory, Center for Cancer ResearchNational Cancer Institute, National Institutes of HealthFrederickUSA
  2. 2.Leidos Biomedical Research Inc., Center for Cancer Research, National Cancer InstituteNational Institutes of HealthFrederickUSA

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