1. CRISPRa induction of Bdnf transcript variants I and IV in primary rat hippocampal neurons.
CRISPRa induction of Bdnf transcript variants I and IV in primary rat hippocampal neurons. A, Bdnf gene structure illustrating non-coding exons (I-IXa) and a common coding exon (IX). sgRNAs were designed upstream of exons I and IV, as indicated by the red and blue lines. B–D, Expression of Bdnf I, IV, and IX transcript variants after targeting dCas9-VPR to exons I and/or IV using sgRNAs, measured with RT-qPCR. B, Bdnf I transcript is specifically upregulated with Bdnf I sgRNA but not with Bdnf IV sgRNA (n = 8, one-way ANOVA, F(3,28) = 15.65, p < ). C, Bdnf IV transcript is specifically upregulated with Bdnf IV sgRNA but not with Bdnf I sgRNA (n = 8, one-way ANOVA, F(3,28) = 34.16, p < ). D, Total Bdnf IX transcript levels are upregulated with both Bdnf I and Bdnf IV sgRNAs (n = 8, one-way ANOVA, F(3,28) = 277.7, p < ). sgRNA designed for the bacterial LacZ gene is used as a non-targeting control in panels B–D. Dunnett’s post hoc test was used for individual comparisons. All data are expressed as mean ± SEM. Individual comparisons; **p < 0.01, ***p < 0.001, ****p <
Katherine E. Savell et al. eNeuro 2019;6:ENEURO
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