title: Gene silencing approaches in mast cells and primary human basophils.
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.
authors.affiliation: Medway School of Pharmacy, The University of Kent, Anson Building, Central Avenue, Chatham Maritime, Kent, ME4 4TB, UK, V.Sumbayev@kent.ac.uk.
authors.forename: Vadim V
authors.initials: VV
authors.lastname: Sumbayev
authors.forename: Bernhard F
authors.initials: BF
authors.lastname: Gibbs
mesh_terms: D001491:Basophils / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry; D006801:Humans; D008407:Mast Cells / Q000166:cytology / Q000378:metabolism*; D061251:Primary Cell Culture / Q000379:methods; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D005229:Fatty Acids, Monounsaturated; D000644:Quaternary Ammonium Compounds; D034741:RNA, Small Interfering; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane