GFP Reporters to Monitor Instability and Expression of Expanded CAG/CTG Repeats.
Abstract
Expanded CAG/CTG repeats are genetically unstable and, upon expression, cause neurological and neuromuscular diseases. The molecular mechanisms of repeat instability and expression remain poorly understood despite their importance for the pathogenesis of a family of 14 devastating human diseases. This is in part because conventional assays are tedious and time-consuming. Recently, however, GFP-based reporters have been designed to provide a rapid and reliable means of assessing these parameters. Here we provide protocols for quantifying repeat instability and expression using a GFP-based chromosomal reporter and the newly developed ParB/ANCHOR-mediated Inducible Targeting (PInT) and how to validate the results.
KEYWORDS:
DNA repair; Expanded CAG/CTG repeat diseases; GFP reporters; Gene expression; Genome stability
- PMID: 31586353