Gene Silencing Approaches in Mast Cells and Primary Human Basophils

  • Vadim V. SumbayevEmail author
  • Bernhard F. Gibbs
Part of the Methods in Molecular Biology book series (MIMB, volume 1192)


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.

Key words

Transfection siRNA Lipofection Electroporation Nanofection 


  1. 1.
    Norata GD, Tibolla G, Catapano AL (2013) Gene silencing approaches for the management of dyslipidaemia. Trends Pharmacol Sci 34:198–205PubMedCrossRefGoogle Scholar
  2. 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 311:195–198PubMedCrossRefGoogle Scholar
  3. 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 39:3511–3519PubMedCrossRefGoogle Scholar
  4. 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:e34259PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Wang Z, Rao DD, Senzer N, Nemunaitis J (2011) RNA interference and cancer therapy. Pharm Res 28:2983–2995PubMedCrossRefGoogle Scholar
  6. 6.
    Imai E, Isaka Y (2002) Gene electrotransfer: potential for gene therapy of renal diseases. Kidney Int 61:S37–S41PubMedCrossRefGoogle Scholar
  7. 7.
    Vilarino N, MacGlashan D Jr (2005) Transient transfection of human peripheral blood basophils. J Immunol Methods 296:11–18PubMedCrossRefGoogle Scholar
  8. 8.
    Burke B (2003) Macrophages as novel cellular vehicles for gene therapy. Expert Opin Biol Ther 3:919–924PubMedCrossRefGoogle Scholar
  9. 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 342–348Google Scholar
  10. 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 19:973–983PubMedCrossRefGoogle Scholar
  11. 11.
    Shatrov VA, Sumbayev VV, Zhou J, Bruene B (2003) Oxidized low density lipoprotein triggers HIF-1alpha protein via redox-dependent mechanism. Blood 101:4847–4849PubMedCrossRefGoogle Scholar
  12. 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 27:677–682PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Medway School of PharmacyThe University of KentKentUK

Personalised recommendations