The ASB6 Knockout HT29 Polyclonal Cells product from Ascent Research provides a CRISPR/Cas9-edited polyclonal knockout cell population featuring targeted disruption of the ASB6 gene in the HT29 human colorectal adenocarcinoma cell line. This genomic modification results in a loss-of-function model that enables investigation of ASB6-dependent processes without the need for transient gene silencing. The polyclonal format retains a heterogeneous knockout population, reflecting the genetic diversity inherent in tumor models, and is suitable for a broad range of functional studies where complete gene disruption is required.
The HT29 cell line is an extensively characterized epithelial model originally derived from a 44-year-old female patient with colorectal adenocarcinoma. HT29 cells are widely employed in cancer biology research, particularly for studies of tumor cell proliferation, differentiation, and drug absorption, due to their ability to form polarized monolayers and express intestinal epithelial markers. This adherent cell line exhibits mutations in APC, TP53, and KRAS, which are common in colorectal cancer, making it a relevant host for investigating molecular mechanisms underlying colorectal carcinogenesis and therapeutic response.
ASB6 encodes the ankyrin repeat and SOCS box containing 6 protein, which functions as a substrate recognition component of an Elongin B/C-Cullin 5-Rbx2 E3 ubiquitin ligase complex. In this capacity, ASB6 promotes the ubiquitination and subsequent proteasomal degradation of target proteins such as the adaptor protein APS (SH2B2) and insulin receptor substrate 1 (IRS1). Through these interactions, ASB6 negatively regulates signaling cascades initiated by cytokines and insulin. Upstream, ASB6 expression is responsive to cytokine stimulation and cellular stress signals. Downstream, loss of ASB6 activity leads to stabilization of APS and IRS1, thereby enhancing Akt and MAPK pathway activation in response to insulin receptor engagement. The complex also includes Elongin B, Elongin C, Cullin 5, and Rbx2 as core components, which collectively mediate ubiquitin transfer to substrates.
In the context of HT29 colorectal cancer cells, ASB6 knockout provides a powerful tool for dissecting the role of ubiquitin-mediated regulation of insulin and cytokine signaling in intestinal epithelial biology. The HT29 background is particularly relevant for exploring how altered protein degradation contributes to metabolic reprogramming in colorectal cancer and insulin resistance. Research utilizing these knockout cells can elucidate whether ASB6-dependent modulation of IRS1 and APS impacts cell proliferation, survival, or migration in colorectal adenocarcinoma, and may uncover links between the ubiquitin-proteasome system and oncogenic signaling.
These polyclonal knockout cells are suitable for a variety of experimental applications. Researchers can employ western blotting and co-immunoprecipitation to assess ASB6 expression and its interaction with Elongin B/C and Cullin 5. Ubiquitination assays can be used to monitor the modification status of APS and IRS1. Insulin stimulation followed by phospho-Akt detection enables functional analysis of insulin signaling enhancement. Additionally, RT-qPCR can quantify ASB6 transcript levels, while flow cytometry, migration assays, and drug sensitivity assays allow comprehensive phenotypic profiling. For further technical details or custom inquiries, please contact Ascent Research.
