The H2AZ2 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cell population engineered to disrupt the H2AZ2 gene in the HEK293T embryonic kidney cell background. This polyclonal knockout pool provides a versatile loss-of-function model for investigating the roles of the H2A.Z-2 histone variant in chromatin biology, transcriptional regulation, and genome maintenance. By eliminating H2AZ2 expression, researchers can interrogate how the absence of this variant histone alters nucleosome composition and affects downstream molecular processes. The cells are supplied as a heterogeneous population, reflecting the combined effects of diverse editing events across the polyclonal pool, without isolation of single-cell clones.
The HEK293T host cell line is derived from human embryonic kidney cells transformed with sheared adenovirus type 5 DNA and stably expressing the SV40 large T antigen. This immortalized cell line supports high-level episomal replication of plasmids containing the SV40 origin of replication, leading to robust transient protein expression and efficient transfection. These characteristics make HEK293T a standard platform for biochemical reconstitution, protein?Cprotein interaction studies, and functional genomics applications. The cell line’s rapid growth and ease of manipulation further enhance its utility as a substrate for generating CRISPR-edited knockout populations.
H2AZ2 encodes the histone variant H2A.Z-2, which is deposited into nucleosomes by the SRCAP chromatin remodeling complex predominantly at promoter and enhancer regions. H2A.Z-2 occupancy modulates chromatin accessibility, thereby regulating the binding of transcription factors and the recruitment of RNA polymerase II. The variant histone interacts with additional chromatin modifiers, including the SWI/SNF complex, and functions in concert with histone chaperones to influence transcriptional programs and DNA repair processes. Its expression is regulated in a cell cycle-dependent manner, and H2A.Z-2 integrates into chromatin dynamics that control genes involved in cell proliferation, differentiation, and stress responses. Consequently, disrupting H2AZ2 alters the balance of histone variant exchange and affects the transcription of downstream target genes governed by H2A.Z occupancy.
In the HEK293T cellular context, knockout of H2AZ2 offers a defined system to dissect the specific contributions of the H2A.Z-2 isoform to gene regulation and genome stability, without the confounding factors present in more complex primary cell models. The polyclonal nature of the knockout population allows for the assessment of average functional effects across multiple genetic backgrounds within an otherwise uniform cell line, which is advantageous for studying epigenetic modifiers. This model enables straightforward correlation of H2A.Z-2 loss with changes in chromatin structure, transcriptome-wide alterations, and DNA damage response pathways, all within a cell type that is amenable to a wide range of molecular biology techniques.
Typical research applications for these polyclonal knockout cells include chromatin immunoprecipitation?Cquantitative PCR (ChIP-qPCR) to map histone modifications and variant occupancy, western blotting to quantify protein-level changes, RNA sequencing to profile transcriptomic shifts, and immunofluorescence to visualize nuclear organization. Additional functional assays, such as flow cytometry and luciferase-based reporter assays, can be employed to monitor cell-cycle perturbations and promoter activity. The H2AZ2 Knockout HEK293T Polyclonal Cells are therefore suited for studies in cancer epigenetics, developmental gene regulation, and chromatin remodeling dynamics. For in-depth technical support or inquiries regarding this product, please contact Ascent Research.
