The HCFC1R1 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the HCFC1R1 gene. Generated by CRISPR/Cas9-mediated gene disruption in the HCT 116 colorectal carcinoma cell line, this product consists of a mixed population of edited alleles, providing a heterogeneous knockout model that avoids clonal artifacts and retains genetic diversity. The polyclonal format ensures robust representation of knockout effects, making it suitable for experiments requiring broad functional assessment rather than single-clone phenotypes. As a non-monoclonal population, these cells are not characterized as biallelic or homozygous knockouts, and researchers should anticipate variable knockout efficiencies across the population. This design is optimal for studying scaffold protein function in cancer signaling, where pathway redundancy and cellular heterogeneity are key considerations.
The host HCT 116 cell line is a human colorectal carcinoma epithelial model derived from a male patient with colon adenocarcinoma. It carries a KRAS G13D mutation and exhibits microsatellite instability (MSI-H), features that mimic aggressive colorectal cancer subtypes. HCT 116 is widely employed in cancer research to investigate oncogenic signaling, drug response, and metastasis, particularly in the context of constitutively active KRAS. Its adherent epithelial morphology and robust growth characteristics facilitate reproducible in vitro experiments, while its tumorigenicity supports xenograft studies. The line??s well-documented molecular landscape makes it an ideal background for dissecting the contributions of scaffold proteins like HCFC1R1 to colorectal cancer pathobiology.
HCFC1R1 (HPIP) functions as a scaffold protein that bridges HCFC1 and PBX1 transcription factors, regulating genes involved in cell cycle progression and epithelial-mesenchymal transition (EMT). It is activated downstream of EGF, TGF-??, and integrin receptors, with oncogenic KRAS strongly amplifying its signaling. HCFC1R1 directly interacts with the p85?? regulatory subunit of PI3K, ERK1/2 kinases, and ??-catenin, thereby coupling upstream signals to PI3K/AKT, MAPK/ERK, and NF-??B pathways. Its downstream targets include cyclin D1 (proliferation), MMP9 (invasion), c-Myc and Bcl-2 (survival), and Snail (EMT). This positions HCFC1R1 as a key integrator of growth factor and stress signals, driving transcriptional programs that promote tumor growth and metastasis.
In the HCT 116 background, where KRAS is constitutively active, HCFC1R1 disruption enables dissection of KRAS-dependent versus -independent signaling outputs. The polyclonal knockout model helps clarify whether HCFC1R1??s effects are predominantly driven by KRAS-mediated PI3K/AKT and MAPK/ERK activation or by alternative inputs like TGF-??/Smad2/3 or integrin/??-catenin. This is especially informative given the MSI-H phenotype, which is associated with distinct immune and therapeutic profiles. Loss of HCFC1R1 in this context can reveal its role in sustaining colorectal cancer cell proliferation, survival, and invasive capacity, offering insights into scaffold-targeted therapeutic strategies.
These polyclonal knockout cells are validated for a range of functional assays, including western blotting and RT-qPCR to confirm gene disruption and assess downstream effectors, proliferation and soft agar colony formation assays to measure tumorigenic potential, and Transwell migration/invasion assays to evaluate metastatic behavior. Co-immunoprecipitation studies can probe HCFC1R1 interactions with HCFC1, PBX1, p85??, and ERK1/2, while phospho-protein analysis and reporter gene assays enable mapping of signaling pathway alterations. The cells are also suitable for xenograft tumor models to examine HCFC1R1??s role in vivo. For further technical details and purchasing information, please contact Ascent Research.
