The APTX Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the aprataxin (APTX) gene has been disrupted via CRISPR/Cas9-mediated gene editing. This loss-of-function model eliminates functional APTX protein expression, enabling researchers to dissect APTX-dependent DNA repair pathways in a human cellular context. As a polyclonal population, this product does not represent a single clonal isolate but rather a heterogeneous pool of edited cells, reflecting the diversity of editing outcomes typical of polyclonal knockout pools.
HEK293T cells are human embryonic kidney epithelial cells that stably express the SV40 large T antigen, which facilitates high-efficiency transient transfection and episomal replication of plasmids containing SV40 origin sequences. This host cell line is widely employed as a versatile platform for protein expression, viral production, and functional genomic studies. Its robust growth characteristics and amenability to genetic manipulation make it an ideal background for generating knockout models to investigate gene function in a human cell-based system.
Aprataxin (APTX) is a DNA repair enzyme that specifically catalyzes the removal of AMP from abortive DNA ligation intermediates, a critical step in the resolution of DNA single-strand breaks. APTX functions downstream of oxidative stress and DNA-damaging agents such as H?O? and ionizing radiation, and its activity is regulated by PARP1 activation. It interacts with core base excision repair and single-strand break repair components, including XRCC1, DNA ligase III, PNKP, and TDP1, to facilitate the repair of 5′-AMP-DNA adducts and reduce DNA strand breaks. Disruption of APTX impairs the repair of oxidative DNA single-strand breaks by failing to remove abortive ligation intermediates, leading to persistent DNA damage and hypersensitivity to genotoxic stress. This molecular pathology underpins ataxia with oculomotor apraxia type 1 (AOA1), an autosomal recessive cerebellar ataxia characterized by neurodegeneration.
In the HEK293T background, APTX knockout results in a deficiency in the repair of oxidative DNA damage, sensitizing the cells to DNA-damaging agents such as hydrogen peroxide, camptothecin, and methyl methanesulfonate. This model recapitulates the DNA repair deficiency observed in AOA1 patient cells and provides a tractable system for studying the molecular consequences of aprataxin loss, including the accumulation of DNA strand breaks and activation of DNA damage response pathways, in a cell line amenable to high-throughput screening and detailed biochemical analyses.
This polyclonal APTX knockout cell population is ideally suited for a variety of research applications, including mechanistic studies of DNA single-strand break repair, modeling of AOA1-associated neurodegeneration, and screening of small molecules that modulate DNA repair pathways. Researchers can utilize these cells in assays such as Western blotting or RT-qPCR to confirm APTX loss, immunofluorescence for ??-H2AX foci to monitor DNA damage, comet assays to measure DNA strand breaks, and cell viability assays following treatment with genotoxic agents. For further information or to place an order, please contact Ascent Research.
