PROKARYOTIC TEST SYSTEM FOR EVALUATION OF OLIGONUCLEOTIDE-AFFECTED ANTISENSE INHIBITION

A new test system is introduced for the estimation of the antisense ef fect of various types of oligodeoxynucleotides. This system is based u pon the inhibition of translation of P2 phage repressor protein. Evide nce for a direct correlation between phage yield and antisense inhibit ion was obtained by comparing the concentration-dependent effectivenes s of an antisense phosphorothioate 18-mer, its mutated version, and th e unmodified oligonucleotide. Oligonucleotides of complementary sequen ces were synthesized against the ribosome binding site and different s ites of the coding region. In our test system only small differences w ere found based on the location of the target sequence. Rather, the de gree of inhibition was related to the calculated ability to hybridize based on T-m values. In this way the sensitivity of the test system co uld also be demonstrated. (C) 1994 Academic Press, Inc.