The ASCC2 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ASCC2 gene in the HeLa human cervical carcinoma cell line. This product provides a loss-of-function model for investigating the roles of ASCC2 in DNA alkylation repair and transcriptional coactivation. By ablating ASCC2 expression, researchers can dissect the gene??s contribution to the DNA damage response and gene regulatory networks, enabling mechanistic studies and drug discovery efforts.
The host HeLa cell line is an immortalized epithelial cervical adenocarcinoma model derived from a human patient. It harbors integrated human papillomavirus type 18 (HPV-18) E6 and E7 oncogenes, which inactivate p53 and retinoblastoma protein (pRb), respectively, driving uncontrolled proliferation. HeLa cells are a cornerstone of cancer research, widely employed for exploring oncogenic mechanisms, genomic instability, and therapeutic resistance. Their robust growth and well-characterized genome make them ideal for generating knockout populations to study tumor biology and DNA repair processes in a relevant cellular context.
ASCC2 (Activating Signal Cointegrator 1 Complex Subunit 2) is a core component of the ASC-1 complex, which also comprises ASCC1, ASCC3, and TRIP4. Mechanistically, ASCC2 acts as a scaffold that recruits the alkylation repair dioxygenase ALKBH3 to sites of DNA alkylation damage, facilitating error-free repair. This function is activated by DNA damage signaling kinases, such as ATM and ATR, and is modulated by p53. Additionally, ASCC2 functions as a transcriptional coactivator by bridging sequence-specific transcription factors with the RNA polymerase II machinery, thereby influencing the expression of downstream target genes involved in cellular stress responses. Thus, ASCC2 integrates DNA repair and transcriptional regulation pathways, with its disruption impacting both genome maintenance and gene expression programs.
In the HeLa cervical cancer model, ASCC2 knockout is particularly relevant for exploring how alkylation damage repair contributes to drug resistance and genomic instability. HPV-driven transformation leads to p53 and pRb inactivation, altering DNA damage checkpoint control and potentially sensitizing or desensitizing cells to alkylating agents. By eliminating ASCC2 function, researchers can assess the dependence of cervical cancer cells on this repair pathway, identify synthetic lethal interactions, and evaluate the ASC-1 complex as a therapeutic target. The polyclonal nature of the knockout population accounts for clonal variation, providing a more representative model for population-level responses.
This product is suitable for a wide range of experimental approaches, including western blotting to confirm ASCC2 loss, immunofluorescence to assess ALKBH3 localization, co-immunoprecipitation to probe ASC-1 complex integrity, and comet assays to measure DNA damage accumulation. Cell viability and colony formation assays enable analysis of response to alkylating chemotherapeutics, while RT-qPCR can monitor transcriptional changes in downstream targets. High-throughput screening of DNA damage response modulators is also facilitated by this knockout model. For additional information or custom orders, please contact Ascent Research.
