Targeted reactivation of FMR1 transcription in FXS embryonic stem cells. 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. Overall design: 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.