Figure 5.

Partial reactivation of FMRP using a dCas9‐Tet1 construct reduces increased spontaneous activity. (a) Immunofluorescent images from FXS neurons (left panel), C1_2 neurons (middle panel) and FXS neurons 2 weeks post‐transduction of 1 MOI dCas9‐Tet1. Blue—nuclei, green—β‐Tubulin and red—FMRP. (b) Quantification of FMRP intensity for C1_2 neurons and FXS neurons transfected with 5 MOI dCas9‐dTet1, 0.5 MOI or 1 MOI dCas9‐Tet1 shows increase FMRP intensity with 1 MOI dCas9‐Tet1. Data normalized to C1_2. (c) Quantification of spontaneous activity recorded on MEAs two weeks post‐transduction demonstrates reduced spontaneous activity from FXS cultures with greater than 5% of C1_2 FMRP expression. Error bars represent ± SEM [Colour figure can be viewed at wileyonlinelibrary.com]