Targeted reactivation of FMR1 transcription in FXS embryonic stem cells

The aim of this work is to selectively increase transcription from the FMR1 locus in FXS hESC lines using CRISPR-Cas9 fused to transcriptional activators. Endonuclease deficient Cas9 (dCas9) was fused to VP192 transcriptional activation domain and transfected in WT and FXS hESCs along with a guide RNA targeted to the CGG repeat. Total RNA was isolated from untransfected WT(UM 4-6), FXS( UM 139-2) hESCs treated with CGG guide RNA and dCas9-VP192 as well as a scramble guide RNA control and dCas9-VP192. RNA seq analysis was carried out on all these samples.