The HCFC1R1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, designed for targeted disruption of the HCFC1R1 gene. This polyclonal pool provides a genetically diverse loss-of-function model that maintains the heterogeneity of the original cells, avoiding the biases associated with clonal selection. Such a population-based knockout approach is particularly advantageous for studies where cellular variability reflects the native tumor microenvironment.
HT29 is a widely characterized epithelial colorectal adenocarcinoma line originally isolated from a primary tumor of a 44-year-old female. The cells harbor truncating mutations in the APC tumor suppressor and TP53, which lead to constitutive activation of the Wnt pathway and compromised DNA damage responses. These features render HT29 a robust model for investigating colorectal cancer biology, including signaling pathway alterations, epithelial-mesenchymal transition, and sensitivity to targeted therapies.
HCFC1R1 encodes a scaffold protein that coordinates signaling downstream of receptor tyrosine kinases and TGF-?? family receptors. It directly interacts with the transcriptional cofactor HCF-1 and the homeodomain transcription factor PBX1, forming complexes that regulate gene expression. Upon stimulation by EGF, TGF-??1, or insulin, HCFC1R1 engages the MAPK/ERK and PI3K/AKT cascades via binding to ERK and AKT, while also associating with SMAD4 to modulate TGF-??/SMAD signaling. This integration results in transcriptional activation of proliferation-promoting genes such as c-Myc and cyclin D1, and contributes to the induction of epithelial-mesenchymal transition through cooperation with SMAD2/3.
In the HT29 colorectal adenocarcinoma context, disruption of HCFC1R1 allows researchers to dissect its functional contributions within an environment of aberrant Wnt signaling and defective p53. This model is instrumental for studying how scaffold proteins like HCFC1R1 sustain mitogenic and survival signals, and for uncovering dependencies that may be exploited for therapeutic intervention. Furthermore, the polyclonal knockout mirrors the heterogeneity of tumor cell populations, making it suitable for studying drug resistance and adaptive signaling rewiring.
This knockout product supports a broad spectrum of experimental assays. Western blotting and immunofluorescence can be used to quantify changes in phospho-ERK and phospho-AKT, while co-immunoprecipitation confirms disrupted interactions with HCF-1 and PBX1. RT-qPCR allows measurement of downstream transcriptional targets such as c-Myc. Cell proliferation can be evaluated via MTT or BrdU incorporation, and migration/invasion potential assessed by transwell assays. Global gene expression profiling by RNA-seq provides comprehensive insights into pathway alterations. For further product information, please contact Ascent Research.
