Monitoring for Epigenetic Modifications at the FMR1 Locus

Silvina Epsztejn-Litman; Rachel Eiges

doi:10.1007/978-1-4939-9080-1_3

Monitoring for Epigenetic Modifications at the FMR1 Locus

Methods Mol Biol. 2019:1942:29-48. doi: 10.1007/978-1-4939-9080-1_3.

Authors

Silvina Epsztejn-Litman¹, Rachel Eiges^{2

3}

Affiliations

¹ Stem Cell Research Laboratory, Medical Genetics Institute, Shaare Zedek Medical Center, Jerusalem, Israel.
² Stem Cell Research Laboratory, Medical Genetics Institute, Shaare Zedek Medical Center, Jerusalem, Israel. rachela@szmc.org.il.
³ Hebrew University Medical School, Jerusalem, Israel. rachela@szmc.org.il.

PMID: 30900173
DOI: 10.1007/978-1-4939-9080-1_3

Abstract

The vast majority of fragile X affected patients do not transcribe FMR1 due to a CGG repeat expansion in the 5'-untranslated region of the FMR1 gene. When the CGGs considerably expand, it elicits abnormal DNA methylation and histone modifications, which are responsible for FMR1 transcriptional silencing. In this chapter, we describe in detail two commonly used protocols for monitoring the epigenetic state of the FMR1 gene that bypass the difficulty in directly analyzing the CGGs. One protocol is for accurately measuring DNA methylation levels and the other is for profiling histone modifications.

Keywords: Bisulfite DNA sequencing; Chromatin immune-precipitation (ChIP); DNA methylation; FMR1; Histone modifications; PCR; qPCR.

MeSH terms

5' Untranslated Regions
DNA Methylation*
Epigenesis, Genetic*
Fragile X Mental Retardation Protein / genetics*
Fragile X Syndrome / diagnosis*
Fragile X Syndrome / genetics*
Gene Silencing
Humans
Trinucleotide Repeats*

Substances

5' Untranslated Regions
FMR1 protein, human
Fragile X Mental Retardation Protein