The H2AJ Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the H2AJ gene has been disrupted to abolish expression of the histone variant H2A.J. This heterogeneous cell pool provides a loss-of-function model for investigating H2A.J-driven chromatin dynamics and inflammatory gene regulation. The polyclonal nature ensures broad mutational representation while avoiding clonal artifacts.
The host cell line, HEK293T, is a human embryonic kidney epithelial derivative that stably expresses the SV40 large T-antigen. Derived from HEK293 cells transformed with adenovirus 5 DNA, it is widely employed for high-level recombinant protein expression and viral packaging due to episomal plasmid amplification. Its epithelial origin and robust growth characteristics make it suitable for a variety of signaling and cytotoxicity studies.
H2A.J functions as a histone H2A variant that accumulates in senescent cells and promotes chromatin accessibility at pro-inflammatory gene promoters. In response to DNA damage or oncogenic stress, the ATM-p53-p21 signaling cascade stabilizes H2A.J, which subsequently interacts with SWI/SNF chromatin remodeling complexes and DNA repair proteins such as KU70/KU80. These interactions drive NF-??B activation, leading to transcriptional upregulation of SASP cytokines like IL-6 and IL-8. H2A.J also coordinates with other histone variants, including H2AX and macroH2A, to orchestrate chromatin reorganization during the DNA damage response.
In HEK293T cells, which can be induced to undergo senescence by genotoxic agents or oncogenic signals, H2AJ knockout eliminates a central mediator of SASP gene expression. This model permits precise dissection of H2A.J-specific effects on chromatin decondensation and NF-??B-dependent cytokine secretion, without interference from endogenous H2A.J. Comparative analysis of wild-type and H2AJ-disrupted cells under stress conditions helps clarify the role of H2A.J in tumor microenvironment modulation and age-related inflammatory disorders.
These cells are suited for senescence-associated ??-galactosidase assays, western blotting and RT-qPCR for H2A.J and SASP markers (IL-6, IL-8), ChIP-qPCR for H2AJ chromatin occupancy, and immunofluorescence for ??-H2AX foci. Additional readouts include flow cytometric cell cycle analysis and RNA-sequencing for global SASP transcriptome profiling. The polyclonal format supports population-scale screening campaigns. For technical support and ordering information, please contact Ascent Research.
