il-13 gene translation is arrested by a novel oligonucleotide in cultured human b- lymphocytes
نویسندگان
چکیده
antisense oligodeoxynucleotides (oligos) are the tools that bind to complemen¬tary sequence of targeted mrna and block specifically protein translation. in the present study, a novel 20 mer oligo as an antisense for human il-13 is introduced. this oligo is designed according to the il-13 mrna coding region and synthesized in two hplc purified and fitc conjugated forms. fluorescence oligo cell uptake is confirmed using flowcytometry and confocal microscopy, and cytotoxicity evaluation is performed us¬ing brdu proliferation assay. human tonsilar b-lymphocytes are purified by positive selection using magnetic cell sorting method and cultured with anti cd40 monoclonal antibody plus ril-4 to induce il-13 production. il-13 antisense is added to medium and real time pcr for mrna, and elisa for protein assays. data indicate that antisense application leads to down regulation and complete suppression of il-13 pro¬tein with no significant effects on mrna, suggesting in vitro protein translation arrest. since 11-13 is a crucial cytokine in allergic conditions, we conclude that interference with the protein synthesis by a nontoxic and efficient antisense oligo can provide an available tool for the investigators on allergic diseases.
منابع مشابه
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عنوان ژورنال:
iranian journal of allergy, asthma and immunologyجلد ۲، شماره ۳، صفحات ۱۳۱-۱۳۷
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