The ASCC2 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-mediated polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line. This product features a disrupted ASCC2 gene, providing a loss-of-function model to investigate the roles of the activating signal cointegrator 1 complex subunit 2 in DNA damage response and genome maintenance. As a heterogeneous polyclonal pool, these cells offer a robust and flexible system for studying population-level effects of ASCC2 depletion without clonal selection artifacts.
HT29 cells are a well-established human colorectal adenocarcinoma epithelial line, known for their mucin-producing capacity and utility as a model of intestinal epithelial biology. These cells form polarized monolayers and retain key characteristics of the colon epithelium, including expression of differentiation markers under specific culture conditions. Their epithelial origin makes them particularly suitable for investigating DNA repair pathways within the context of intestinal cell physiology, a tissue frequently exposed to genotoxic agents from dietary and microbial sources.
ASCC2 functions as a core component of the ASC-1 complex (ASCC), which also includes ASCC1, ASCC3, and TRIP4. This complex is activated by DNA damage stimuli, including UV radiation and alkylating agents, downstream of the ATR kinase. ASCC2 interacts with ALKBH3, a demethylase that removes alkylation lesions from DNA, and the ubiquitin ligases RNF113A and CUL4A, facilitating ubiquitin-proteasome-mediated signaling. Through these interactions, ASCC2 coordinates transcription-coupled nucleotide excision repair and resolution of transcription-blocking lesions, ensuring RNA polymerase II restart and preventing genome instability. The ASCC complex thus integrates signals from the ATM/ATR kinases with post-translational modification machineries to maintain transcriptional integrity.
In HT29 colorectal adenocarcinoma cells, ASCC2 knockout provides a relevant platform to dissect how intestinal epithelial cells handle alkylation damage, which is particularly important given the colon’s exposure to dietary carcinogens and chemotherapeutic alkylating agents. Disruption of ASCC2 in this mucin-producing epithelial background allows researchers to explore links between DNA repair deficiencies and colorectal cancer susceptibility, as well as to model neurodevelopmental defects associated with congenital bone fractures, conditions linked to ASCC2 mutations. The knockout model may reveal how compromised genome maintenance contributes to epithelial barrier dysfunction or malignant transformation.
Researchers can employ the ASCC2 Knockout HT29 Polyclonal Cells in a variety of assays, including western blotting to confirm loss of ASCC2 protein, immunofluorescence staining for ??H2AX to monitor DNA double-strand breaks, and clonogenic survival assays following treatment with methyl methanesulfonate (MMS) to assess sensitivity to alkylating agents. Comet assays provide a measure of DNA strand breaks, while RT-qPCR and RNA-seq enable transcriptomic analysis of DNA damage response pathways. These cells are ideally suited for siRNA or drug library screens, investigation of cisplatin or temozolomide resistance mechanisms, and studies of transcription-coupled repair dynamics. For additional product information or to discuss custom applications, please contact Ascent Research.
