Targeted Reactivation of FMR1 Transcription in Fragile X Syndrome Embryonic Stem Cells

Frontiers in Molecular Neuroscience
Jill M HaenflerP K Todd

Abstract

Fragile X Syndrome (FXS) is the most common inherited cause of intellectual disability and autism. It results from expansion of a CGG nucleotide repeat in the 5' untranslated region (UTR) of FMR1. Large expansions elicit repeat and promoter hyper-methylation, heterochromatin formation, FMR1 transcriptional silencing and loss of the Fragile X protein, FMRP. Efforts aimed at correcting the sequelae resultant from FMRP loss have thus far proven insufficient, perhaps because of FMRP's pleiotropic functions. As the repeats do not disrupt the FMRP coding sequence, reactivation of endogenous FMR1 gene expression could correct the proximal event in FXS pathogenesis. Here we utilize the Clustered Regularly Interspaced Palindromic Repeats/deficient CRISPR associated protein 9 (CRISPR/dCas9) system to selectively re-activate transcription from the silenced FMR1 locus. Fusion of the transcriptional activator VP192 to dCas9 robustly enhances FMR1 transcription and increases FMRP levels when targeted directly to the CGG repeat in human cells. Using a previously uncharacterized FXS human embryonic stem cell (hESC) line which acquires transcriptional silencing with serial passaging, we achieved locus-specific transcriptional re-activation of F...Continue Reading

References

Aug 1, 1992·Nature Genetics·A K GedeonG R Sutherland
Jul 15, 1994·American Journal of Medical Genetics·S L NolinC S Dobkin
Feb 1, 1997·Nature Genetics·H E MalterB A Oostra
Feb 28, 1998·Human Molecular Genetics·P ChiurazziG Neri
Nov 9, 2000·Molecular Diagnosis : a Journal Devoted to the Understanding of Human Disease Through the Clinical Application of Molecular Biology·B GoldC S Chiang
Jul 14, 2001·American Journal of Medical Genetics·F TassoneP J Hagerman
Sep 17, 2002·American Journal of Human Genetics·Bradford CoffeeDaniel Reines
Oct 23, 2003·Human Molecular Genetics·Stephanie CemanStephen T Warren
Jun 29, 2004·Trends in Neurosciences·Mark F BearStephen T Warren
Nov 26, 2004·Human Molecular Genetics·Roberta PietrobonoGiovanni Neri
Dec 21, 2007·Neuron·Gül DölenMark F Bear
Mar 11, 2008·Nature Chemical Biology·Shuang ChangStephen T Warren
Oct 22, 2008·Molecular and Cellular Biology·Miri KimStephanie Ceman
Nov 8, 2008·Bioinformatics·Adi Laurentiu TarcaRoberto Romero
Mar 6, 2009·Genome Biology·Ben LangmeadSteven L Salzberg
Mar 18, 2009·Bioinformatics·Cole TrapnellSteven L Salzberg
Oct 25, 2011·Annual Review of Pathology·Michael R SantoroStephen T Warren
Feb 24, 2012·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Vijayalaxmi C NalavadiGary J Bassell
Apr 10, 2012·Annual Review of Neuroscience·Asha L BhakarMark F Bear
Apr 17, 2012·Neuron·Aubin MichalonLothar Lindemann
Dec 12, 2012·Nature Biotechnology·Cole TrapnellLior Pachter
Jul 23, 2013·Nature Biotechnology·Patrick D HsuFeng Zhang
Jul 31, 2013·Nature Methods·Pablo Perez-PineraCharles A Gersbach
Oct 15, 2013·American Journal of Medical Genetics. Part a·Elisabetta Tabolacci, Pietro Chiurazzi

❮ Previous
Next ❯

Citations

Nov 5, 2019·Current Pharmaceutical Design·Michael Telias
Feb 16, 2020·The Journal of Biological Chemistry·Alexandra N Khristich, Sergei M Mirkin
Jul 31, 2019·International Journal of Molecular Sciences·Renée H L RaaijmakersDerick G Wansink
Sep 4, 2020·Journal of Neurochemistry·Thomas J TurnerGabriele Lignani
Jan 24, 2019·Brain Sciences·Carolyn M Yrigollen, Beverly L Davidson
Mar 14, 2019·Brain Sciences·Rakhi Pal, Aditi Bhattacharya
Feb 19, 2020·Nature Neuroscience·Caitlin M RodriguezPeter K Todd
Aug 28, 2020·Frontiers in Bioengineering and Biotechnology·Ksenia M ShakirovaErdem B Dashinimaev
May 4, 2020·Journal of Neurodevelopmental Disorders·Khaleel A RazakCraig A Erickson
Jan 7, 2021·International Journal of Molecular Sciences·Nasir JavaidSangdun Choi
Jan 29, 2021·The Journal of Biological Chemistry·Alexandra N Khristich, Sergei M Mirkin
Apr 25, 2021·Acta Neuropathologica Communications·M Rebecca GlineburgPeter K Todd
May 1, 2021·International Journal of Molecular Sciences·Tayma Handal, Rachel Eiges
Jun 1, 2021·Frontiers in Cellular Neuroscience·Keqin XuPeng Jin

❮ Previous
Next ❯

Methods Mentioned

BETA
Transfection
fluorescence
electrophoresis
transfections
PCR
RNA-seq

Software Mentioned

Cufflinks
Bowtie2
CuffDiff
FastQC
TopHat
iPathwayGuide
Tuxedo Suite
CummeRbund R package

Related Concepts

Related Feeds

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

Autism

Autism spectrum disorder is associated with challenges with social skills, repetitive behaviors, and often accompanied by sensory sensitivities and medical issues. Here is the latest research on autism.