Analysis of Trinucleotide Repeat Stability by Integration at a Chromosomal Ectopic Site.

We describe a general strategy for the construction of clonal cell lines containing CNG repeats of various lengths, in which the microsatellites are integrated using the yeast FLP recombinase at a single ectopic recombination acceptor site in the HeLa genome. We illustrate two types of (CTG/CAG) cell lines, one of which contains dual fluorescent marker genes flanking the (CTG/CAG) repeat, and one which does not. We show that long CNG repeats are prone to DNA double strand breaks (DSBs) upon exposure of these cell lines to prolonged replication stress.
PMID: 31586345 [PubMed - in process]

Source: Mol Biol Cell - October 7, 2019 Category: Molecular Biology Authors: Gadgil RY, Rider SD, Lewis T, Barthelemy J, Leffak M Tags: Methods Mol Biol Source Type: research