The IFI27 Knockout HCT 116 Polyclonal Cells product constitutes a CRISPR/Cas9-edited polyclonal knockout cell population in which the IFI27 gene has been disrupted in the HCT 116 human colorectal carcinoma cell line. This loss-of-function model enables researchers to interrogate the cellular and molecular consequences of IFI27 depletion without reliance on transient silencing methods, offering a stable genetic background for long-term studies. As a polyclonal pool, the population preserves genetic heterogeneity while eliminating functional IFI27 protein, making it suitable for experiments where clonal variability is undesirable. The product facilitates investigation of interferon-mediated apoptosis, innate immune responses, and cell cycle control in an epithelial colorectal cancer context.
The host cell line, HCT 116, is a widely used epithelial cell model derived from human colorectal carcinoma. It harbors a KRAS G13D mutation, which activates downstream oncogenic signaling, while retaining a microsatellite stable (MSS) genomic profile and wild-type p53 status. These characteristics render HCT 116 particularly relevant for studying colorectal cancer biology, including tumor suppression mechanisms, apoptotic pathways, and responses to cytokine stimulation. The adherent, epithelial morphology and robust growth kinetics further support reproducible experimental workflows, including drug treatment, viral infection, and time-series analyses.
IFI27, also known as ISG12a, functions as an interferon-stimulated gene product that localizes to the mitochondrial intermembrane space. Its expression is potently induced by type I interferons (IFN-??/??) through the JAK-STAT signaling cascade, involving the receptors IFNAR1 and IFNAR2, kinases JAK1 and TYK2, and the transcription factor complex STAT1/STAT2/IRF9; additional transcriptional regulation is mediated by IRF1. Once expressed, IFI27 interacts with BCL2 family proteins and BAX to promote mitochondrial outer membrane permeabilization, leading to the release of apoptogenic factors, activation of caspase-9 and caspase-3, and execution of apoptosis. Beyond its pro-apoptotic role, IFI27 negatively regulates viral replication, likely by modulating mitochondrial innate immune platforms, and influences cell cycle progression through modulation of cyclin D1 levels.
In the HCT 116 colorectal cancer background, IFI27 knockout provides a powerful tool to dissect the interplay between interferon signaling and tumor cell survival. Given the KRAS G13D mutation, which enhances proliferative and anti-apoptotic pathways, the absence of IFI27 may alter the balance between apoptosis and resistance to interferon-induced cell death. This model is valuable for exploring how mitochondrial apoptotic priming affects colorectal carcinoma cell fate, particularly in the context of innate immune stimulation. Moreover, because HCT 116 cells are susceptible to viral infection, the knockout population enables mechanistic studies of IFI27-dependent antiviral restriction without confounding host genetic variability.
This polyclonal knockout cell population supports a broad range of experimental applications. Researchers can employ Western blotting and RT-qPCR to confirm IFI27 disruption and quantify downstream targets such as caspase-3, BAX, and cyclin D1. Flow cytometry and Annexin V staining facilitate apoptosis assessment following interferon stimulation, while co-immunoprecipitation assays enable interrogation of IFI27 interactions with BCL2 family members and BAX. Cell viability assays and viral replication studies offer complementary functional readouts. The model is well-suited for drug discovery efforts targeting the interferon-JAK-STAT apoptotic axis in colorectal cancer and for basic investigations of innate immune defense. For additional details, please contact Ascent Research.
