The IPP Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population in which the IPP gene has been disrupted. This product provides a heterogeneous pool of HCT 116 cells with targeted IPP gene disruptions, created via CRISPR/Cas9 genome editing. The polyclonal format avoids clonal selection bias and is well-suited for pooled functional assays. This loss-of-function model enables in-depth study of IPP’s role in ubiquitin-mediated proteostasis and cancer cell physiology.
HCT 116 is a human colorectal carcinoma epithelial cell line derived from a male patient. It harbors a KRAS G13D mutation and a CTNNB1 (??-catenin) stabilizing mutation, accompanied by microsatellite instability (MSI). These genetic features drive constitutive RAS/MAPK and Wnt/??-catenin signaling, making HCT 116 a standard model for colorectal tumorigenesis. This background is ideal for evaluating gene knockouts that intersect with oncogenic pathways.
IPP encodes a BTB domain-containing substrate adaptor for the Cullin-3?CRING E3 ubiquitin ligase complex. It directly interacts with CUL3 and RBX1 to recruit substrates for ubiquitination by E2 enzymes and subsequent proteasomal degradation. Although IPP’s specific substrates are unknown, it is predicted to target proteins controlling cell cycle progression or stress responses. IPP expression may be regulated by cell cycle transcription factors, though upstream control remains poorly defined.
In HCT 116 cells, IPP knockout likely disrupts degradation of key substrates, potentially leading to their accumulation and dysregulation of protein homeostasis. This may alter the balance of oncogenic signaling networks driven by KRAS and ??-catenin, affecting proliferation, survival, or proteotoxic sensitivity. Thus, this model is valuable for investigating how CUL3-mediated proteolysis impacts colorectal cancer cell biology.
These polyclonal knockout cells can be used in co-immunoprecipitation to probe CUL3 complex assembly, cycloheximide chase assays to assess protein stability, and ubiquitination assays to monitor polyubiquitin changes. Functional studies include cell proliferation, colony formation, and survival analyses. Western blotting and RT-qPCR confirm IPP disruption and downstream effects. This resource facilitates identification of IPP substrates and elucidates the contribution of the Cullin-3 E3 ligase to colorectal cancer pathogenesis. For further information, contact Ascent Research.
