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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Verkerk AJ, Pieretti M, Sutcliffe JS, Fu YH, Kuhl DP, Pizzuti A et al (1991) Identification of a gene (FMR-1) containing a CGG repeat coincident with a breakpoint cluster region exhibiting length variation in fragile X syndrome. Cell 65:905–914. https://doi.org/10.1016/0092-8674(91)90397-h
Oberlé I, Rousseau F, Heitz D, Kretz C, Devys D, Hanauer A et al (1991) Instability of a 550-base pair DNA segment and abnormal methylation in fragile X syndrome. Science 252:1097–1102. https://doi.org/10.1126/science.252
Hayward BE, Kumari D, Usdin K (2017) Recent advances in assays for the fragile X-related disorders. Hum Genet 136(10):1313–1327. Review
Coffee B, Zhang F, Ceman S, Warren ST, Reines D (2002) Histone modifications depict an aberrantly heterochromatinized FMR1 gene in fragile x syndrome. Am J Hum Genet 71:923–932
Coffee B, Zhang F, Warren ST, Reines D (1999) Acetylated histones are associated with FMR1 in normal but not fragile X-syndrome cells. Nat Genet 22:98–101
Pietrobono R, Tabolacci E, Zalfa F, Zito I, Terracciano A, Moscato U (2005) Molecular dissection of the events leading to inactivation of the FMR1 gene. Hum Mol Genet 14:267–277
Tabolacci E, Moscato U, Zalfa F, Bagni C, Chiurazzi P, Neri G (2008) Epigenetic analysis reveals a euchromatic configuration in the FMR1 unmethylated full mutations. Eur J Hum Genet 16:1487–1498
Tabolacci E, Pietrobono R, Moscato U, Oostra BA, Chiurazzi P, Neri G (2005) Differential epigenetic modifications in the FMR1 gene of the fragile X syndrome after reactivating pharmacological treatments. Eur J Hum Genet 13:641–648
Kumari D, Usdin K (2010) The distribution of repressive histone modifications on silenced FMR1 alleles provides clues to the mechanism of gene silencing in fragile X syndrome. Hum Mol Genet 19(23):4634–4642
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Epsztejn-Litman, S., Eiges, R. (2019). Monitoring for Epigenetic Modifications at the FMR1 Locus. In: Ben-Yosef, D., Mayshar, Y. (eds) Fragile-X Syndrome. Methods in Molecular Biology, vol 1942. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9080-1_3
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9080-1_3
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9079-5
Online ISBN: 978-1-4939-9080-1
eBook Packages: Springer Protocols