Triplex Targeting of a Native Gene in Permeabilized Intact Cells: Covalent Modification of the Gene for the Chemokine Receptor CCR5

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1998 Apr 4

PMID 9469844

Citations 13

Authors

E S Belousov

I A Afonina

I V Kutyavin

A A Gall

M W Reed

H B Gamper

R M Wydro

R B MEYER

Affiliations

Soon will be listed here.

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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