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. 1998 Mar 1;26(5):1324–1328. doi: 10.1093/nar/26.5.1324

Triplex targeting of a native gene in permeabilized intact cells: covalent modification of the gene for the chemokine receptor CCR5.

E S Belousov 1, I A Afonina 1, I V Kutyavin 1, A A Gall 1, M W Reed 1, H B Gamper 1, R M Wydro 1, R B Meyer 1
PMCID: PMC147384  PMID: 9469844

Abstract

A 12 nucleotide oligodeoxyribopurine tract in the gene for the chemokine receptor CCR5 has been targeted and covalently modified in intact cells by a 12mer triplex forming oligonucleotide (TFO) bearing a reactive group. A nitrogen mustard placed on the 5'-end of the purine motif TFO modified a guanine on the DNA target with high efficiency and selectivity. A new use of a guanine analog in these TFOs significantly enhanced triplex formation and efficiency of modification, as did the use of the triplex-stabilizing intercalator coralyne. This site-directed modification of a native chromosomal gene in intact human cells under conditions where many limitations of triplex formation have been partially addressed underscores the potential of this approach for gene control via site-directed mutagenesis.

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Selected References

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