Experimental Controls

Antisense experiments should be specifically targeted against a chosen complementary sequence. Anionic phosphorothioate-containing oligonucleotides and cationic agents used to increase their uptake may be toxic in a nonspecific manner. Hence, for specificity tests, it is necessary to use dose-dependent controls of cells treated only with cationic agents and those using the same cationic agent plus a scrambled control phosphorothioate-containing oligonucleotide with the same base composition but in an order different from that chosen for the putatively active AS-ODN. This scrambled control oligonucleotide should be inactive and lacking homology to any other gene. Moreover, the active AS-ODN should primarily reduce expression of the complementary mRNA and preferentially block the corresponding pathway controlled by expression of this gene (11,12).

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