RNAi-mediated inhibition of tumor necrosis factor alpha-related conditions
A condition, the technology of the subject, applied in the direction of DNA / RNA fragments, recombinant DNA technology, medical preparations containing active ingredients, etc., can solve the problem of not teaching the target sequence, etc.
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[0151] Example 1 is used to specifically silence the interfering RNA of TNFR1 in GTM-3 cells
[0152] This study examined the ability of TNFR1 interfering RNA to knock down endogenous TNFR1 protein expression levels in cultured GTM-3 cells.
[0153] Use TNFR1 siRNAs, siCONTROL RISC-free-siRNA #1, or siCONTROL Non-Targeting siRNA #2 (NTC2) and Standard in vitro concentrations (0.1-10 nM) of #1 transfection reagent (Dharmacon, Lafayette, CO) accomplished transfection of GTM-3 cells (Pang, I.H. et al., 1994. Curr. Eye Res. 13:51-63). Dissolve all siRNA in 1X siRNA buffer, which is 20mM KCl, 6mM HEPES (pH7.5), 0.2mM MgCl 2 of aqueous solution. Control samples included a buffer control in which the volume of siRNA was replaced with an equal volume of IX siRNA buffer (-siRNA). Western blotting was performed using an anti-TNFR antibody (Santa Cruz Biotechnology, Santa Cruz, CA) to assess TNFRl protein expression. TNFR1 siRNA is a double-stranded interfering RNA specific for the...
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