The ATF2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population designed for disruption of the human ATF2 gene in HAP1 cells. This knockout model provides a heterogeneous pool of gene-edited cells generated through CRISPR/Cas9-mediated targeting, enabling loss-of-function studies without clonal isolation. The polyclonal format maintains genetic diversity, which can enhance the robustness of experimental findings in cell-based assays.
The HAP1 cell line is a near-haploid human cell model derived from the KBM-7 chronic myeloid leukemia line. These cells exhibit a fibroblastoid morphology and adhere to plastic surfaces in culture. They are male and retain BCR-ABL expression, reflecting their leukemic origin. The near-haploid karyotype reduces the likelihood of functional complementation by wild-type alleles, facilitating efficient gene disruption in polyclonal knockout populations and making HAP1 an ideal host for genetic studies.
ATF2 encodes a basic leucine zipper transcription factor that integrates stress signals via the JNK and p38 MAPK pathways. Upstream, small GTPases RAC1 and CDC42 signal through MAP3K1 to MAP2K4 (MKK4) and MAP2K7 (MKK7), which activate JNK1 and JNK2; alternatively, MAP2K3 (MKK3) and MAP2K6 (MKK6) activate p38??. Phosphorylation by these kinases promotes ATF2 dimerization with partners such as c-Jun and CREB1, leading to transactivation of target genes containing CRE/AP-1 response elements. Representative downstream targets include CCND1, JUN, TNF, IFNB1, CDKN1A, and BCL2L11. ATF2 also interacts with transcriptional coactivators EP300 and CREBBP, as well as SUMO1, to orchestrate programs governing proliferation, apoptosis, and development.
In the HAP1 background, knockout of ATF2 abolishes stress-induced transcriptional responses, creating a rigorous loss-of-function model for dissecting kinase signaling cascades. The BCR-ABL-positive context is particularly relevant for leukemia research, enabling studies of oncogenic stress adaptation and drug resistance. This model supports investigation of ATF2’s interactions with cofactors such as EP300 and CREBBP, and assessment of downstream effectors like BCL2L11 in apoptotic regulation. The polyclonal nature reflects tumor heterogeneity, enhancing utility for drug screening and genetic interaction mapping.
Researchers can apply this product in diverse experimental workflows, including western blotting for total and phospho-ATF2, RT-qPCR analysis of transcriptional targets, and luciferase reporter assays to measure ATF2 activity. The cells are well-suited for apoptosis assays using Annexin V/PI staining, cell viability tests with MTS, and drug sensitivity screens for melanoma, breast cancer, and leukemia compounds. Chromatin immunoprecipitation (ChIP) enables profiling of ATF2 genomic occupancy. For further technical information or to place an order, please contact Ascent Research.
