Skip to main content

Gene Silencing Approaches in Mast Cells and Primary Human Basophils

  • Protocol
  • First Online:
Basophils and Mast Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1192))

Abstract

The ability to silence gene expression is an invaluable tool for elucidating the importance of intracellular signaling proteins which contribute to the effector functions of mast cells and basophils. However, primary mast cells and their terminally differentiated blood counterpart, namely basophils, pose a difficult challenge for gene silencing approaches given not only their state of maturation and difficulty to transfect, but also because their functions are readily altered by cell-handling conditions. Here, we describe a method using lipofection which has been successfully employed to silence gene expression using siRNA in human LAD2 mast cells as well as primary human basophils.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Norata GD, Tibolla G, Catapano AL (2013) Gene silencing approaches for the management of dyslipidaemia. Trends Pharmacol Sci 34:198–205

    Article  CAS  PubMed  Google Scholar 

  2. Macrae I, Zhou K, Li F, Repic A, Brooks A, Cande W, Adams P, Doudna J (2006) Structural basis for double-stranded RNA processing by dicer. Science 311:195–198

    Article  CAS  PubMed  Google Scholar 

  3. Sumbayev VV, Nicholas SA, Streatfield CL, Gibbs BF (2009) Involvement of hypoxia-Inducible Factor-1 in IgE-mediated primary human basophil responses. Eur J Immunol 39:3511–3519

    Article  CAS  PubMed  Google Scholar 

  4. Sumbayev VV, Yasinska IM, Oniku AE, Streatfield CL, Gibbs BF (2012) Involvement of hypoxia-inducible factor-1 in the inflammatory responses of human LAD2 mast cells and basophils. PLoS One 7:e34259

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Wang Z, Rao DD, Senzer N, Nemunaitis J (2011) RNA interference and cancer therapy. Pharm Res 28:2983–2995

    Article  CAS  PubMed  Google Scholar 

  6. Imai E, Isaka Y (2002) Gene electrotransfer: potential for gene therapy of renal diseases. Kidney Int 61:S37–S41

    Article  PubMed  Google Scholar 

  7. Vilarino N, MacGlashan D Jr (2005) Transient transfection of human peripheral blood basophils. J Immunol Methods 296:11–18

    Article  CAS  PubMed  Google Scholar 

  8. Burke B (2003) Macrophages as novel cellular vehicles for gene therapy. Expert Opin Biol Ther 3:919–924

    Article  CAS  PubMed  Google Scholar 

  9. Sumbayev VV, Yasinska IM, Gibbs BF (2013) Biomedical applications of gold nanoparticles. In: Said A, Tang CH, Oprisan S (eds) Recent advances in circuits, communications and signal processing, WSEAS Press, Athens, pp 342–348

    Google Scholar 

  10. Nicholas SA, Sumbayev VV (2009) The involvement of hypoxia-inducible factor 1 alpha in Toll-like receptor 7/8-mediated inflammatory response. Cell Res 19:973–983

    Article  CAS  PubMed  Google Scholar 

  11. Shatrov VA, Sumbayev VV, Zhou J, Bruene B (2003) Oxidized low density lipoprotein triggers HIF-1alpha protein via redox-dependent mechanism. Blood 101:4847–4849

    Article  CAS  PubMed  Google Scholar 

  12. Kirshenbaum AS, Akin C, Wu Y, Rottem M, Goff JP et al (2003) Characterization of novel stem cell factor responsive human mast cell lines LAD 1 and 2 established from a patient with mast cell sarcoma/leukemia; activation following aggregation of FcepsilonRI or FcgammaRI. Leuk Res 27:677–682

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Vadim V. Sumbayev .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Sumbayev, V.V., Gibbs, B.F. (2014). Gene Silencing Approaches in Mast Cells and Primary Human Basophils. In: Gibbs, B., Falcone, F. (eds) Basophils and Mast Cells. Methods in Molecular Biology, vol 1192. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1173-8_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1173-8_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1172-1

  • Online ISBN: 978-1-4939-1173-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics