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Cellular Delivery of siRNAs Using Bolaamphiphiles

  • Kshitij GuptaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1632)

Abstract

Discovery of RNA interference (RNAi) has opened up a new arena of therapeutic intervention for the treatment of cancerous as well as noncancerous diseases. The RNAi pathway utilizes RNAi inducers such as small interfering RNAs (siRNAs) to target and silence disease causing genes. However, efficient delivery of siRNAs for eliciting efficacious RNAi has remained a daunting challenge. Nonviral vectors such as lipids have shown great promise in delivering siRNAs. Recently, a novel class of cationic lipid molecules “bolaamphiphile lipids” or “bola lipids” has been shown to deliver siRNAs to cause effective gene silencing in cells. The present chapter showcases the ability of bola lipids to form micelles, bind with nucleic acids and protect nucleic acids against nucleases. Also, high in vitro transfection efficiency for silencing green fluorescent protein (GFP) using Dicer substrate siRNAs (dsiRNAs) designed against GFP at nontoxic dose in a human breast cancer model is demonstrated. Our results showed that these cationic bola lipids are promising siRNA delivery agents.

Key words

Bolaamphiphiles RNAi RNA–DNA hybrids dsiRNAs Green fluorescent protein Human breast cancer cells 

Notes

Acknowledgments

This work 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 No. HHSN261200800001E. This research was supported [in part] by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research. 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. Dr. Kshitij Gupta also acknowledges “Scientist pool officer” position awarded by Council of Scientific and Industrial Research, New Delhi, India and “Young Scientist” award “YSS/2014/000937” by Science and Engineering Research Board, Department of Science & Technology, India.

References

  1. 1.
    Sledz CA, Williams BR (2005) RNA interference in biology and disease. Blood 106(3):787–794CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Elbashir SM, Lendeckel W, Tuschl T (2001) RNA interference is mediated by 21- and 22-nucleotide RNAs. Genes Dev 15(2):188–200CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Afonin KA et al (2015) Triggering of RNA interference with RNA-RNA, RNA-DNA, and DNA-RNA nanoparticles. ACS Nano 9(1):251–259CrossRefPubMedGoogle Scholar
  4. 4.
    Liu J et al (2004) Argonaute2 is the catalytic engine of mammalian RNAi. Science 305(5689):1437–1441CrossRefPubMedGoogle Scholar
  5. 5.
    Meister G et al (2004) Human Argonaute2 mediates RNA cleavage targeted by miRNAs and siRNAs. Mol Cell 15(2):185–197CrossRefPubMedGoogle Scholar
  6. 6.
    Valencia-Sanchez MA et al (2006) Control of translation and mRNA degradation by miRNAs and siRNAs. Genes Dev 20(5):515–524CrossRefPubMedGoogle Scholar
  7. 7.
    Whitehead KA, Langer R, Anderson DG (2009) Knocking down barriers: advances in siRNA delivery. Nat Rev Drug Discov 8(2):129–138CrossRefPubMedGoogle Scholar
  8. 8.
    Parlea L et al (2016) Cellular delivery of RNA nanoparticles. ACS Comb Sci 18(9):527–547CrossRefPubMedGoogle Scholar
  9. 9.
    Gupta K et al (2015) Bolaamphiphiles as carriers for siRNA delivery: From chemical syntheses to practical applications. J Control Release 213:142–151CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Zhang S, Zhao Y, Zhi D (2012) Non-viral vectors for the mediation of RNAi. Bioorg Chem 40(1):10–18CrossRefPubMedGoogle Scholar
  11. 11.
    Zhang S, Zhi D, Huang L (2012) Lipid-based vectors for siRNA delivery. J Drug Target 20(9):724–735CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Kanasty R et al (2013) Delivery materials for siRNA therapeutics. Nat Mater 12(11):967–977CrossRefPubMedGoogle Scholar
  13. 13.
    Kim T et al (2013) In silico, in vitro, and in vivo studies indicate the potential use of bolaamphiphiles for therapeutic siRNAs delivery. Mol Ther Nucleic acids 2:e80CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Afonin KA et al (2013) Activation of different split functionalities on re-association of RNA-DNA hybrids. Nat Nanotechnol 8(4):296–304CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Afonin KA et al (2014) Multifunctional RNA nanoparticles. Nano Lett 14(10):5662–5671CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Grinberg S et al (2005) Novel cationic amphiphilic derivatives from vernonia oil: synthesis and self-aggregation into bilayer vesicles, nanoparticles, and DNA complexants. Langmuir 21(17):7638–7645CrossRefPubMedGoogle Scholar
  17. 17.
    Popov M et al (2010) Cationic vesicles from novel bolaamphiphilic compounds. J Liposome Res 20(2):147–159CrossRefPubMedGoogle Scholar
  18. 18.
    Kim DH et al (2005) Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy. Nat Biotechnol 23(2):222–226CrossRefPubMedGoogle Scholar
  19. 19.
    Rose SD et al (2005) Functional polarity is introduced by Dicer processing of short substrate RNAs. Nucleic Acids Res 33(13):4140–4156CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Popov M et al (2013) Delivery of analgesic peptides to the brain by nano-sized bolaamphiphilic vesicles made of monolayer membranes. Eur J Pharm Biopharm 85(3 Pt A):381–389CrossRefPubMedGoogle Scholar
  21. 21.
    Dulbecco R, Freeman G (1959) Plaque production by the polyoma virus. Virology 8(3):396–397CrossRefPubMedGoogle Scholar
  22. 22.
    Bindewald E et al (2016) Multistrand structure prediction of nucleic acid assemblies and design of RNA switches. Nano Lett 16(3):1726–1735CrossRefPubMedGoogle Scholar
  23. 23.
    Afonin KA et al (2011) Design and self-assembly of siRNA-functionalized RNA nanoparticles for use in automated nanomedicine. Nat Protoc 6(12):2022–2034CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.RNA Structure and Design Section, RNA Biology Laboratory, National Cancer InstituteNational Institutes of HealthFrederickUSA
  2. 2.Department of Inorganic and Physical ChemistryIndian Institute of ScienceBangaloreIndia

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