Russian Journal of AllergyRussian Journal of Allergy1810-88302686-682XFarmarus Print Media68310.36691/RJA683Research ArticleSIRNA-MEDIATED INHIBITION OF INTERLEUKINE-13 PRODUCTION IN VITROShilovskiyI P-MazurovD V-ShershakovaN N-KhaitovM Rmkhaitov@immune.umos.ruInstitute of Immunology1512201296242710032020Copyright © 2012, Pharmarus Print Media2012Background. According to current views, one of the major mediators involved in the development of allergic process is IL-13. The goal of this work was to design small interfering RNA molecules to effectively inhibit il-13 gene expression of mice in experiments in vitro. Methods. For the expression of IL-13 in in vitro gene coding sequence il-13 was amplified using cDNA ConA-stimulated spleen cells from BALE / c mice as a template and cloned into the expression vector pUCHR IRES GFP. Using a computer analysis were designed six variants of siRNA, directed against mRNA-il-13. To test the efficiency of siRNA a co-transfection of 1x 105 cells HEK293T mixture (0,5 mg and 1 mg of plasmid siRNA) coupled with Lipofectamine 2000 reagent was carried out. Twenty-four hours later, the gene expression changes in il-13 recorded by flow cytometry on the fluorescence intensity of GFP+-cells. Gene expression of il-13 mRNA was assessed by quantitative PCR, and the level of the protein product by ELISA. results. As a result, siRNA molecules were obtained and three of them were able to effectively inhibit the gene expression of il-13. Conclusion. Thereby variants of siRNA, which can effectively inhibit the production of mice’s IL-13 in vitro; can be used later in experiments in vivo so to understand the role of IL-13 in the pathogenesis of allergic conditions as to develop new therapy approaches.allergyinterleukin-13inhibition of il-13 gene expressionsiRNAmechanism of RNA interferenceаллергияинтерлейкин-13подавление экспрессии гена il-13миРНКмеханизм интерференции РНК[Coyle A.J., Le Gros G., Bertrand C. et al. Interleukin-4 is required for the induction of lung Th2 mucosal immunity. Am. J. Respir. Cell. Mol. Biol. 1995, v. 13, p. 54-59.][Gavett S.H., O’Hearn D.J., Karp C.L. et al. Interleukin-4 receptor blockade prevents airway responses induced by antigen challenge in mice. Am. J. Physiol. 1997, v. 272, p. L253-L261.][Aman M.J., Tayebi N., Obiri N. I. еt al. cDNA cloning and characterization of the human interleukin-13 receptor alpha chain. J. Biol. Chem. 1996, v. 271, p. 29265-29270.][Walter D.M., McIntire J.J., Berry G. et al. Critical role for IL-13 in the development of allergen-induced airway hyperreactivity. J. Immunol. 2001, v. 167, p. 4668-4675.][Zhu Z., Homer R. J., Wang Z. et al. Pulmonary expression of interleukin-13 causes inflammation, mucus hypersecretion, subepithelial fibrosis, physiologic abnormalities, and eotaxin production. J. Clin. Invest. 1999, v. 103, p. 779-788.][Meister G., Tuschl T. Mechanisms of gene silencing by doublestranded RNA. Nature. 2004, v. 431, p. 343-349.]