The ATF1 Knockout HAP1 Polyclonal Cells are a polyclonal knockout cell population derived from the HAP1 human haploid cell line, engineered via CRISPR/Cas9-mediated disruption of the ATF1 gene. This loss-of-function model provides a heterogeneous pool of knockout cells suitable for pooled screening and bulk functional assays. The polyclonal format reflects the inherent genetic variation of CRISPR editing, offering a robust system for investigating ATF1-dependent signaling in a near-haploid genetic background.
HAP1 is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) cell line. Its haploid karyotype greatly facilitates genetic manipulation and functional genomics, including large-scale knockout screens and drug sensitivity profiling. Widely used as a model for hematological malignancies, HAP1 cells are particularly valuable for studying gene function in a simplified genetic context.
ATF1 is a transcription factor that binds cAMP response elements (CRE) and integrates signals from multiple pathways. Upon phosphorylation by upstream kinases such as the PKA catalytic subunit, ERK1/2, and CaMKII??activated by growth factors (EGF, NGF) and cytokines (IL-1, TNF-alpha)??ATF1 recruits coactivators like CBP/p300 to drive target gene expression. Through interactions with CREB1, CREM, ATF2, and JUN, it regulates key targets including CCND1 for proliferation, BCL2 and MCL1 for survival, and FOS, JUN, and NR4A1 for stress responses. ATF1 thus serves as a critical node in cAMP/PKA, MAPK/ERK, PI3K/AKT, and calcium signaling cascades.
In the CML-derived HAP1 background, ATF1 knockout provides a model to dissect its role in leukemogenesis and drug sensitivity. ATF1??s involvement in oncogenic fusions like EWSR1-ATF1 in clear cell sarcoma, as well as in melanoma and neurodegenerative disorders, further broadens the utility of this knockout population. Combined with the haploid genome, it enables efficient genetic interaction mapping and synthetic lethality screens.
This model supports diverse research applications, including transcriptomic profiling by RNA-seq, targeted chromatin analysis by ChIP-qPCR, and CRE-luciferase reporter assays to assess transcriptional activity. Cell proliferation and apoptosis assays characterize growth and survival phenotypes, while drug sensitivity screening identifies pathway dependencies. Immunofluorescence and Western blotting confirm ATF1 disruption and downstream signaling changes. For further information or to discuss custom applications, please contact Ascent Research.
