Figure 8.

CRISPR genome editing in cultured mouse astrocytes. (a) We generated two lentivirus constructs containing the human GFAP promotor, Cas9, sgRNA, and fluorescent reporters EGFP and mCherry. (b,c) We infected cultured mouse astrocytes with lentiviruses encoding CAS9‐EGFP and sgRNAs targeting GFAP (b) and Sox9 (c) with the mCherry reporter. Images were taken 7 days after infection. Arrows point to cells doubly infected with lentiviruses encoding CAS9 and sgRNA. Doubly infected cells had reduced GFAP (b) or Sox9 (c) protein expression (d). Quantification of percentage of GFAP and Sox9 positive cells with or without CRISPR genome editing [Color figure can be viewed at wileyonlinelibrary.com]