The ASB9 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from HT29 human colorectal adenocarcinoma cells, with targeted disruption of the ASB9 gene. This loss-of-function model facilitates investigation of ASB9’s role in ubiquitin-mediated protein degradation and colorectal cancer biology. The polyclonal nature provides a robust system reflecting diverse genomic edits without clonal selection biases.
HT29 is a well-characterized intestinal epithelial cell line isolated from a primary colorectal adenocarcinoma of a 44-year-old female Caucasian. It serves as a key model for colorectal cancer research, retaining properties of aberrant proliferation and differentiation relevant to tumorigenesis and drug response studies.
ASB9 acts as a substrate recognition subunit of the cullin-5-RING E3 ubiquitin ligase complex, interacting with CUL5, ELOB, ELOC, and RBX2 to ubiquitinate target proteins for proteasomal degradation. This complex is regulated upstream by STAT3 transcription factor activity and promoter methylation, integrating cytokine signals with protein turnover. While specific substrates are unknown, they likely include cell cycle and apoptosis regulators, positioning ASB9 in control of proliferation and survival via the ubiquitin-proteasome pathway.
In HT29 colorectal cancer cells, ASB9 knockout enables dissection of its role in proliferation, apoptosis evasion, and drug resistance. Disruption of ASB9-mediated substrate targeting may alter stability of proteins involved in oncogenic signaling, affecting cellular responses to chemotherapy. This model thus aids in understanding how ubiquitination dysregulation promotes colorectal cancer progression and in identifying therapeutic targets within the ubiquitin-proteasome system.
Key applications include mechanistic ubiquitination studies, substrate identification via co-immunoprecipitation and ubiquitination assays, and protein turnover analysis using cycloheximide chase. Functional assessments such as MTT proliferation, Annexin V apoptosis flow cytometry, colony formation, and cell cycle flow cytometry can be performed, along with RT-qPCR for gene expression. These cells are suited for drug sensitivity screening targeting the ubiquitin-proteasome pathway in colorectal cancer. For further details, contact Ascent Research.
