The KAT6A Knockout HEK293T Polyclonal Cells product comprises a heterogeneous population of human embryonic kidney HEK293T cells subjected to CRISPR/Cas9-mediated disruption of the KAT6A gene. This polyclonal knockout pool provides a robust loss-of-function model for investigating KAT6A-dependent epigenetic and transcriptional mechanisms without clonal selection or single-cell isolation. The polyclonal format preserves the inherent biological variability of the knockout event across the cell population, making it suitable for bulk analyses where averaged gene-editing effects are sufficient and cost-effective.
The parental HEK293T cell line is derived from human embryonic kidney epithelium and stably expresses the SV40 large T antigen, which promotes episomal replication of plasmids containing the SV40 origin. This feature renders the cell line exceptionally efficient for transient protein overexpression and recombinant virus production, including lentivirus and adenovirus. HEK293T cells maintain a near-diploid karyotype and are a standard model in molecular and cellular biology laboratories for studying gene function, signal transduction, and cancer-related pathways.
KAT6A (MOZ) is a histone acetyltransferase of the MYST family that acts as a crucial transcriptional coactivator. It acetylates histone H3 at lysine 14 (H3K14ac) and lysine 23 (H3K23ac), relaxing chromatin and facilitating gene activation. KAT6A assembles into multimeric complexes with ING5, BRPF1, MEAF6, and EP300 to modify chromatin landscapes. The acetyltransferase is recruited to target loci by RUNX1 and CBFB, potently activating the HOXA gene cluster essential for developmental patterning and hematopoietic specification. KAT6A also participates in p53 transcriptional activation and intersects with Notch, Wnt, and DNA damage pathways. Its dysregulation via translocations or mutations is linked to acute myeloid leukemia, intellectual disability, and developmental disorders.
In the HEK293T background, the KAT6A knockout provides a genetically defined system to interrogate KAT6A-dependent transcriptional programs and histone acetylation dynamics. Although HEK293T cells are of kidney origin, they express core transcriptional machinery and coactivators relevant to many tissues, and their high transfection efficiency allows complementation with wild-type or mutant KAT6A constructs to map functional domains. This model is particularly valuable for studying epigenetic mechanisms that underlie hematopoietic gene expression and leukemogenesis, as KAT6A??s transactivation functions can be analyzed in an easily manipulable cellular environment.
Researchers can employ these polyclonal knockout cells to explore KAT6A??s role in histone modification dynamics, transcriptional coactivation, and chromatin remodeling. Typical experimental workflows include western blotting to assess global H3K14ac and H3K23ac levels, RT-qPCR to measure expression of HOXA9 and other downstream targets, and ChIP-qPCR to profile KAT6A-dependent acetylation at specific gene promoters. Additionally, luciferase reporter assays can dissect KAT6A??s coactivator function, while colony formation and proliferation assays help delineate its impact on cell growth and survival. These cells are also suitable as a genetic background for epigenetic inhibitor screening. For more information, technical support, or ordering details, please contact Ascent Research.
