The ANP32E Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for investigating the cellular functions of ANP32E, a histone chaperone involved in chromatin remodeling and signal transduction. This product consists of a heterogeneous pool of HEK293T cells harboring CRISPR/Cas9-mediated disruption of the ANP32E gene, enabling loss-of-function studies without clonal isolation. The polyclonal format provides a robust and reproducible model for examining ANP32E-dependent phenotypes while minimizing clonal artifacts, making it suitable for a broad range of experiments where genetic homogeneity is not essential.
HEK293T cells are a widely used human embryonic kidney cell line stably expressing the SV40 large T antigen, which promotes episomal replication of plasmids containing the SV40 origin of replication. This characteristic renders HEK293T cells highly amenable to transient transfection and robust protein expression, establishing them as a preferred host for mechanistic studies, viral production, and signaling assays. The parental line was originally derived by transforming human embryonic kidney cells with sheared adenovirus 5 DNA, resulting in a well-characterized model that supports high-level recombinant protein production and efficient genetic manipulation.
ANP32E functions as a histone chaperone that specifically facilitates the deposition of the histone variant H2A.Z into chromatin, a process critical for transcriptional regulation, DNA repair, and genome stability. Through its interaction with the SRCAP chromatin remodeling complex, ANP32E modulates nucleosome dynamics at gene promoters. Additionally, ANP32E binds and inhibits the catalytic subunit of protein phosphatase 2A (PP2A), leading to sustained activation of downstream kinases such as AKT and ERK1/2. This dual role links chromatin architecture to proliferative signaling cascades. Upstream, ANP32E expression is transcriptionally regulated by E2F1 and MYC, placing it within a network that coordinates cell cycle progression and apoptotic responses. ANP32E also associates with CRM1, suggesting involvement in nucleocytoplasmic transport.
In the HEK293T background, disruption of ANP32E is expected to impair H2A.Z incorporation at target loci, altering the transcriptional landscape and potentially sensitizing cells to PP2A-mediated dephosphorylation of AKT and other substrates. Given the robust expression of SV40 large T antigen, which itself perturbs cell cycle control, the knockout model may reveal context-specific dependencies on ANP32E for proliferation and survival. This system is particularly valuable for dissecting the interplay between chromatin remodeling and oncogenic signaling, as well as for understanding how ANP32E contributes to viral pathogenesis, since HEK293T cells are frequently used in viral replication studies.
Researchers can employ this polyclonal knockout pool in a variety of assays to interrogate ANP32E biology. Western blotting and co-immunoprecipitation are suitable for assessing ANP32E protein levels and its interactions with H2A.Z or the PP2A catalytic subunit. Chromatin immunoprecipitation followed by quantitative PCR (ChIP-qPCR) allows mapping of H2A.Z occupancy changes at specific promoters. PP2A activity assays and flow cytometry for apoptosis can elucidate signaling and cell death phenotypes. Transcriptome-wide analyses via RNA-seq and functional viral replication assays further expand the utility of this model. For more information, please contact Ascent Research. For additional technical details, please contact Ascent Research.
