CRIPSR/Cas9 SAMs-directed activation of PrP expression.

(A) sqRT PCR evaluation of Prnp SAM 1 and 2; “*”—p<0.001; Kruskal-Wallis test. (B) Immunoblot (SAF83) assessment of PrP levels after SAM activation using sgRNAs #1 and #2. (C) FACS of N2a cells transfected with Prnp-directed SAMs; histograms shows increased mean fluorescence intensity (MFI) corresponding to cell surface PrP (SAF83) on cells transfected with Prnp SAM 1 and 2 (dotted and solid line, respectively) as compared with MFI of “no_sgRNA2.0” control (in grey). (D) Immunostaining of N2a cells transfected with Prnp-directed SAMs; PrP levels on cells transfected with functional SAMs were increased as compared to untransfected (EGFP negative) cells in the same FOV. In case of “no sgRNA2.0” control, no differences between transfected and untransfected cells with respect to PrP staining were observed.