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RNA interference (RNAi),a posttranscriptional gene silencing process mediated by small double-stranded RNA specifically complementary to the targeted transcript,has been used extensively in the developmentof novel therapeutic approaches against various human diseases including chronic myelogenous leukemia (CML).Here.we report the successful construction of a tetracycline-controlled siRNA in CML cell line K562.A K562cell line stably expressing the reverse tetracycline-controlled transactivator (rtTA) was constructed.A tetracy-cline responsive element(TRE) was integrated into the RNA polymerase Ⅲ promoter region of pBS/U6 thatwas used to drive specific siRNA to target the novel cytokine receptor-like factor 3 (CRLF3) gene,The resultsshow that rtTA was able to recognize the TRE to prevent siRNA-mediated exogenous and endogenous CRLF3gene repressions.Moreover,CRLF3-siRNA mediated gene repression could be induced in a dose-dependentmanner in the presence of doxycycline.Thus,the inducible siRNAi system in K562 cells might be useful for thestudy of RNAi-mediated therapeutic approaches against CML.
RNA interference (RNAi), a posttranscriptional gene silencing process mediated by small double-stranded RNA specifically complementary to the targeted transcript, has been used extensively in the development of novel therapeutic approaches against various human diseases including chronic myelogenous leukemia (CML). Here.we report the successful construction of a tetracycline-controlled siRNA in CML cell line K562. A K562cell line stably expressing the reverse tetracycline-controlled transactivator (rtTA) was constructed. A tetracy-cline responsive element (TRE) was integrated into the RNA polymerase III promoter region of pBS / U6 that was used to drive specific siRNA to target the novel cytokine receptor-like factor 3 (CRLF3) gene, The resultsshow that rtTA was able to recognize the TRE to prevent siRNA-mediated exogenous and endogenous CRLF3gene repressions. Moreover, CRLF3 -siRNA mediated gene repression could be induced in a dose-dependentmanner in the presence of doxycycline.Thus, the inducible siRNAi system in K562 cells might be useful for the study of RNAi-mediated therapeutic approaches against CML.