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Communication Dans Un Congrès Année : 2017

In vitro studies of the long-range regulation of FOXL2 expression by epigenome editing

Résumé

It has been shown that Forkhead boxL2 gene (FOXL2) is a master gene for female gonadal development. Firstly, it is involved in ovarian determination[1] and/or maintenance[2], and secondly it is necessary for ovarian follicle growth[3]. Interestingly, long-range deletions affecting FOXL2 expression have been reported in different species. In goats, a natural mutation called PIS for “Polled Intersex Syndrome” is responsible for female-to-male sex-reversal in homozygous XX PIS-/- mutant animals. This mutation corresponds to a 11.7kb homozygous deletion[4] lying at around 300 kb upstream of FOXL2 gene[5,6]. The PIS mutation is responsible for FOXL2 expression silencing and the sex-reversal[1]. In humans, various heterozygous long-range deletions have been shown to be responsible for decreasing FOXL2 expression, resulting in Blepharophimosis Ptosis Epicanthus inversus Syndrome[7]. To date, the Smallest Region of deletion Overlap (SRO) described in human patients, is a 7.4 kb region, encompassing the long non-coding RNA PISRT1 that was shown, in the goat species, to be closely regulated with FOXL2. All this data suggest that this distant region located at around 300 kb of FOXL2 may contain crucial regulatory elements for its expression. Analyzes of the epigenetic state of this region in human ovary (ChIP-Seq, ENCODE data), or of its counterpart in mouse ovary (ChIP-qPCR, personal data) revealed H3K27ac enrichment, which is an epigenetic mark preferentially observed on active gene enhancer or promoter. In order to prove that this region contains crucial enhancer(s), but also to demonstrate that long-range epigenetic modifications are able to control FOXL2 expression, we developed a program of epigenome editing. The CRISPR/dCas9 technology has been previously deflected to induce epigenetic modifications at specific loci[8]. Thus we designed several guide RNAs, coupled to a CRISPR/dCas9-p300 which is capable to acetylate H3K27. In a cellular model, different regions will be targeted, focusing on the most conserved elements of the SRO counterpart.
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Dates et versions

hal-01605214 , version 1 (02-10-2017)

Identifiants

  • HAL Id : hal-01605214 , version 1
  • PRODINRA : 395760

Citer

Clara Gobe, Nathalie Daniel-Carlier, Geneviève Jolivet, Marjolaine André, Eric Pailhoux, et al.. In vitro studies of the long-range regulation of FOXL2 expression by epigenome editing. 3. Journée de Séminaires du Département Phase sur l'Epigénétique EpiPhase, May 2017, Jouy-en-Josas, France. 1 p. ⟨hal-01605214⟩
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