Supplementary Figure 1: CRISPRi shows prominent targeting specificity in neurons.
From: CRISPR interference-based specific and efficient gene inactivation in the brain
(a, e and i) Design of Syt1- and Snap25-targeting sgRNA variant arrays to test the binding of dCas9-KRAB to these two ‘false’ targets. The colored nucleic acids indicate mismatches. (b, f and j) ChIP-qPCR analysis of dCas9-KRAB binding to the targeting sites with the navigation of sgRNA variants (n = 3 for all groups). (c, g and k) qRT-PCR analysis of mRNA expression in neurons expressing different sgRNA variants (n = 3 for all groups). (d, h and l) Immunoblot analysis and quantification of protein expression in neurons with co-expression of sgRNA variants and dCas9-KRAB (n = 3 for all groups). In d, h and l, blots were cropped; full-length blots are presented in Supplementary Figure 9. All data were obtained from neurons from three independent cultures with similar results and shown as mean ± s.e.m. Statistical significance was assessed by Unpaired Student’s t-test. *P < 0.05. For detailed numbers and statistical analysis, see Supplementary Table 3.
