Silencing of Bruton's tyrosine kinase (Btk) using short interfering RNA duplexes (siRNA)

FEBS Letters

Heinonen, Juhana E.¹  Nore, Beston F.¹  Smith, C.I.Edvard¹ 

[1] Clinical Research Center (CRC), KFC-Novum, Karolinska Institutet, Huddinge University Hospital, SE-141 86 Huddinge, Sweden

关键词: Immunodeficiency;    X-linked agammaglobulinemia;    Mast cell;    Allergy;    Btk;    Tec;    Bmx;    siRNA;    RNAi;    Btk;    Bruton's tyrosine kinase;    FcϵRI;    high-affinity IgE receptor of mast cells;    LFM-A13;    α-cyano-β-hydroxy-β-methyl-N-(2;    5-dibromophenyl) propenamide;    EGFP;    enhanced green fluorescent protein;    BCR;    B-cell receptor;   

DOI  :  10.1016/S0014-5793(02)03206-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Tec family tyrosine kinases, Bruton's tyrosine kinase (Btk), Itk, Bmx, Tec, and Txk, are multi-domain proteins involved in hematopoietic signaling. Here, we demonstrate that human Btk protein can transiently be depleted using double-stranded short RNA interference (siRNA) oligonucleotides. Imaging and Western blotting analysis demonstrate that Btk expression is down regulated in heterologous systems as well as in hematopoietic lineages, following transfection or microinjection of Btk siRNA duplexes. The induction of histamine release, a pro-inflammatory mediator, in RBL-2H3 mast cells was reduced by 20–25% upon Btk down regulation. Similar, results were obtained when the Btk activity was inhibited using the kinase blocker LFM-A13. These results demonstrate a direct role of Btk for the efficient secretion of histamine in allergic responses.

【 授权许可】

Unknown