Induced recombination mediated by pyrimidine TFOs

Copyright information:

Taken from "Triplex-induced recombination and repair in the pyrimidine motif"

Nucleic Acids Research 2005;33(11):3492-3502.

Published online 16 Jun 2005

PMCID:PMC1151591.

© The Author 2005. Published by Oxford University Press. All rights reserved

() Schematic representation of the pSupFTC18 vector. This SV40-based vector contains two mutant genes in tandem. The upstream mutant gene, , contains a C-to-T point mutation at nucleotide position 163. The downstream mutant gene, , contains a G-to-A point mutation at nucleotide position 115. In between the two mutant genes is an 18 bp A–T-rich homopurine/homopyrimidine target site. The chemically modified oligonucleotides described in were designed to bind to this target sequence. () Triplex-induced recombination in COS-7 cells. The plasmid described in () was pre-transfected into COS-7 cells followed 1 day later by transfection of the indicated oligonucleotides or mock transfection. After 48 h, shuttle vector DNA was isolated and transformed into indicator bacteria to analyze gene function and allow quantification of recombination events. Percent recombination was calculated by the ratio of the number of blue colonies to total colonies.