The ATF1 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal human cell population engineered for functional knockout of the ATF1 gene. Derived from the SK-HEP-1 hepatocellular carcinoma cell line, this mixed polyclonal pool is generated via CRISPR-mediated gene disruption, providing a versatile model to study ATF1-dependent signaling while preserving the genetic heterogeneity inherent to tumor cell populations.
The SK-HEP-1 host cell line is an epithelial, adherent cell type originally derived from ascites of a liver adenocarcinoma patient. It serves as a standard in vitro model for hepatocellular carcinoma research, widely used for investigating oncogenic pathways, drug resistance, and cancer cell biology in the context of hepatic malignancies.
ATF1 is a basic leucine zipper transcription factor that binds cAMP response elements (CRE) and is activated by phosphorylation at Ser63 downstream of cAMP/PKA and MAPK (ERK1/2, p38) signaling. Upon phosphorylation, ATF1 recruits the coactivators CBP/p300 and forms homo- or heterodimers with CREB1, ATF2, or ATF3 to drive transcription of target genes such as c-FOS, CCND1, BCL2, BDNF, and NR4A1. ATF1 thereby integrates signals from GPCR?Cadenylate cyclase?CcAMP?CPKA cascades and receptor tyrosine kinase?CRAS?CRAF?CMEK?CERK?CRSK/MSK pathways to regulate proliferation, survival, and stress adaptation.
In hepatocellular carcinoma, ATF1 contributes to malignant phenotypes by promoting G1/S progression via cyclin D1 and blocking apoptosis through BCL2 induction. Upstream kinases like PKA, ERK1/2, and p38 MAPK are frequently dysregulated in HCC, leading to aberrant ATF1 activity that may modulate sensitivity to therapeutic agents such as sorafenib and cisplatin. Knockout of ATF1 in SK-HEP-1 cells thus enables dissection of its role in liver cancer cell growth, migration, and drug response, providing a clinically relevant loss-of-function model.
The knockout cells support a broad array of assays: Western blot for total and phospho-ATF1; RT-qPCR or RNA-seq for transcriptional changes in c-FOS, CCND1, and other targets; ChIP-qPCR to quantify ATF1 occupancy at CRE; and luciferase reporter assays for cAMP responsiveness. Functional studies can include MTT proliferation, Transwell migration, and drug sensitivity analyses with sorafenib or cisplatin. Applications span investigation of ATF1 in HCC, cAMP?CMAPK crosstalk, and screening of CREB/ATF pathway inhibitors. For additional information, please contact Ascent Research.
