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. Author manuscript; available in PMC: 2021 Nov 1.
Published in final edited form as: Nature. 2020 Oct 21;587(7833):281–284. doi: 10.1038/s41586-020-2835-2

Figure 1. Screen to identify SpCas9 gRNAs that unsilence paternal Ube3a.

Figure 1.

a. Paternal Ube3a-ATS locus showing the location of genes, transcription collision region (dashed lines), and gRNA library target sites.

b. Percentage of Ube3am+/pYFP neurons co-stained for UBE3A-YFP and tdTomato; co-transfected with CamKIIα promoter:tdTomato, SpCas9 and one gRNA (circles) or treated with 300 nM topotecan for 72 h. Black circles, P<0.005 and fold change >4.

c. Ube3am+/pYFP neurons co-transfected with CamKIIα promoter:tdTomato, SpCas9 and negative control gRNA (neg.) or Spjw33 gRNA. Nuclei, blue (DAPI). Scale bar, 100 μm.

d. Expression of the indicated genes seven days after transducing Ube3am+/pYFP cortical neurons with lentiviral SpCas9+Spjw33, relative to SpCas9+neg. control gRNA (dashed line). qPCR, normalized to Eif4a2, n=3. Error bars, standard error of the mean (S.E.M.), * P<0.01.