Fig. 3: AAV2/1-dNmCas9 exhibits dose-dependent effects in vitro.
From: AAV-dCas9 vector unsilences paternal Ube3a in neurons by impeding Ube3a-ATS transcription
a Representative images of mouse primary mouse cortical neurons transduced by AAV2/1-dNmCas9 and immunostained for NmCas9 and NeuN. Scale bar, 10 µm. b The percentage of transfected Ube3am+/pYFP mouse neurons positive for UBE3A-YFP following transfection with the indicated Cas9 variant and gRNA. Number of Snord115 targets and strand (t template, n non-template) are indicated. n = 4 culture wells per condition. Error bars indicate SEM. c–f Relative expression of (c) patUbe3a, (d) Ube3a-ATS, (e) Snord115 snoRNAs, and (f) NmCas9 in neurons following dosing with AAV2/1-dNmCas9-Nmg15 or NmCas9-Nmg15 at the viral multiplicity of infection (MOI) indicated. Normalized to Eif4a2, n = 2–3 wells per condition per dose. Expression normalized to non-targeting gRNA virus (dashed line). Error bars indicate SEM. [inhibitor] or [agonist] vs normalized response non-linear regression analysis was used for curve fitting.
