The HERC5 Knockout HT29 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population in which the HERC5 gene has been disrupted, generating a loss-of-function model for investigating HERC5-dependent cellular mechanisms. This polyclonal cell product is derived from the HT29 host cell line through targeted gene disruption, enabling pooled knockout populations that preserve heterogeneous genetic backgrounds and avoid clonal artifacts. The product is well-suited for studying HERC5-related roles in antiviral innate immunity, ubiquitin-like signaling, and inflammatory regulation without the constraints of monoclonal selection.
The HT29 cell line is a well-characterized human colorectal adenocarcinoma epithelial cell line originally established from a primary tumor. It serves as a widely accepted in vitro model for intestinal epithelial barrier function and colorectal cancer pathophysiology. HT29 cells retain key epithelial features, including tight junction competence and differentiation potential, making them invaluable for dissecting epithelial cell biology, tumor progression, and therapeutic responses in a relevant colorectal cancer context.
HERC5 encodes an E3 ubiquitin ligase that catalyzes ISG15 conjugation (ISGylation), a critical post-translational modification in antiviral defense. HERC5 expression is robustly induced by type I interferon (IFN-??/??) and IFN-?? signaling through the canonical JAK-STAT pathway: upon IFN receptor engagement, JAK1 and TYK2 phosphorylate STAT1 and STAT2, which dimerize and recruit IRF9 to form the ISGF3 transcription factor complex that drives transcription of ISG15 and HERC5. HERC5 then acts in concert with the E1 activating enzyme UBA7 and the E2 conjugating enzyme UBE2L6 (UbcH8) to conjugate ISG15 onto lysine residues of target proteins, including viral proteins and host antiviral effectors such as MxA, PKR, and RIG-I. Additionally, HERC5-mediated ISGylation can modulate the NF-??B pathway by targeting signaling components, thereby influencing inflammation and apoptosis. Key interacting partners include ISG15, UBE2L6, and UBA7, while upstream regulators include NF-??B and IRF9.
In the HT29 colorectal adenocarcinoma model, HERC5 knockout enables precise dissection of how interferon-driven ISGylation impacts tumor cell-intrinsic behaviors. Colorectal cancers are frequently subjected to inflammatory microenvironments where interferon and NF-??B signaling shape tumor growth, immune evasion, and response to therapy. By removing HERC5, researchers can interrogate its contribution to ISG15 conjugate formation, alterations in NF-??B transcriptional activity, and downstream effects on apoptosis and migration in an epithelial tumor context, providing insights into colorectal cancer biology and potential vulnerabilities.
This HERC5 knockout polyclonal cell product supports a broad range of experimental applications, including Western blotting analysis of HERC5 protein levels and ISG15 conjugation patterns, RT-qPCR and RNA-seq for transcriptional profiling, immunofluorescence and flow cytometry to assess pathway activation, antiviral infection assays to evaluate innate immune competence, NF-??B luciferase reporter assays for signaling readouts, and apoptosis or migration assays to probe functional outcomes. It is an essential tool for researchers studying antiviral innate immunity, ISGylation dynamics, interferon signaling networks, and the intersection of ubiquitin-like modifications with colorectal cancer pathogenesis. For more information or to inquire about this product, please contact Ascent Research.
