The H2BC17 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population providing a loss-of-function model for the H2BC17 gene. This polyclonal pool avoids clonal selection, offering a heterogeneous population ideal for studying H2BC17 deficiency effects. Derived from the HEK293T host cell line, it serves as a versatile tool for chromatin biology research.
HEK293T cells, a human embryonic kidney epithelial line transformed with adenovirus 5 DNA, stably express the SV40 large T-antigen. This enables episomal plasmid replication and high-level transient protein expression, supporting applications such as lentivirus production and transient transfection. As a widely used model for kidney cell biology, HEK293T provides a robust and easily manipulated platform for CRISPR/Cas9-mediated gene disruption.
H2BC17 encodes a core histone H2B variant that is a crucial nucleosome component, forming octamers with H2A, H3, and H4 to package DNA and regulate transcription, replication, and repair. Its expression is controlled by E2F transcription factors and Cyclin E/CDK2, and it is phosphorylated by ATM/ATR kinases upon DNA damage. H2BC17 interacts with histone chaperones NAP1 and FACT, chromatin assembly factor CAF-1, and acts upstream of transcription factors SP1 and NF-??B, as well as DNA repair protein BRCA1. Knockout of H2BC17 disrupts nucleosome assembly, causing chromatin destabilization, genome instability, and aberrant gene expression.
In the HEK293T context, H2BC17 knockout enables dissection of histone variant functions in a widely used cell line. The polyclonal nature reflects population heterogeneity, which is advantageous for studying varied genetic responses. This model is particularly suited for investigating DNA damage repair pathways and chromatin dynamics, given HEK293T cells’ common use in genomic stability research. Additionally, the presence of SV40 large T-antigen may intersect with chromatin pathways, offering insights into viral oncoprotein effects on histone biology.
This knockout cell population supports diverse assays, including ChIP for nucleosome occupancy, immunofluorescence for chromatin organization, and RNA-seq for transcriptional changes. Western blotting assesses histone levels, while cell cycle analysis and ??H2AX foci detection probe DNA damage responses. Apoptosis assays further evaluate cellular outcomes. These make it valuable for epigenetics, cancer biology, and cell cycle studies. For further information, please contact Ascent Research.
