Fig. 1

The generation of CREB1 knockout (KO) cells and rescue cells. A The generation of CREB1 knockout cells by CRISPR/Cas9 technology. A pair of gRNAs were designed to target the upstream and downstream sequences of CREB1-exon 1. After puromycin selection, CREB1 knockout clones carrying homozygous deletion of exon 1 were screened out by PCR. Clone 1–3, pool (all clones mixture), and WT cells were verified by Sanger sequencing. P–F and P–R indicate the locations of the targeted sequence by gRNA. B The generation of CREB1 rescued cells. Lenti CREB1 Hygro plasmid was constructed by cloning CREB1 cDNA and EF1α-driven HygroR into LentiCRISPRv2 plasmid. Lenti CREB1 Hygro was co-transfected with three packaging plasmids into HEK293T cells. Virus particles were collected and used to infect 3 different CREB1 knockout clones. Finally, CREB1 rescued cells were obtained after hygromycin selection. C. Western blot of indicated proteins in wild type (WT), CREB1 KO, and CREB1 rescue cells