Fig. 2

The CRISPR/Cas9 induces mutations in Creb. a Agarose gel electrophoresis shows the result of the Surveyor assay in the targeted Creb loci of control Neuro2a cells (control) and the cloned CRISPR/Cas9 vector transfected cells (CRISPR/Cas9). Plus and minus signs above each lane indicate the presence and absence of T7 endonuclease I, respectively. An arrowhead indicates the PCR product (655 bp). Two arrows indicate the digested fragments of the PCR product by T7 endonuclease I. b Representative mutation patterns revealed by DNA sequencing of the target site in the exon 7 of Creb. Top indicates the targeted sequence (blue underline) and PAM sequence (red underline) in Creb. An arrowhead indicates the Cas9 cut site. Red dots indicate deleted bases. Red characters indicate base substitutions. The number of deletions (−) and base substitutions (S) are shown. c Western blot analysis was performed with anti-CREB and β-actin antibodies. The lysates were prepared form controls (control) and the cloned transfected cells (CRISPR/Cas9)