The DTX3L Knockout HGC-27 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HGC-27 human gastric adenocarcinoma cell line, featuring disruption of the DTX3L gene. DTX3L encodes an E3 ubiquitin ligase critical for ubiquitin-dependent signaling in DNA damage repair and innate immunity. The polyclonal format provides a heterogeneous pool of loss-of-function alleles, suitable for pooled functional studies without clonal selection.
The HGC-27 cell line originates from a metastatic gastric adenocarcinoma and carries a TP53 mutation that impairs p53-mediated tumor suppression. This genetic context makes the cells particularly useful for modeling advanced gastric cancer, where genomic instability and aberrant signaling drive progression. HGC-27 is a well-established model for investigating oncogenic pathways, drug responses, and tumor microenvironment interactions.
DTX3L is an E3 ubiquitin ligase that forms a complex with PARP9 to ubiquitinate substrates in the DNA damage response and interferon pathways. Activated by IFN-?? and type I interferons via JAK-STAT signaling, DTX3L enhances STAT1 transcriptional activity and modulates NF-??B signaling. It also ubiquitinates histone H2B and interacts with PARP14 and DTX family members. Knockout of DTX3L disrupts these ubiquitin-dependent processes, impairing cellular responses to genotoxic stress and immune stimuli. This model allows dissection of the interplay among STAT1, NF-??B, and the ubiquitin-proteasome system in gastric cancer.
In the TP53-mutant HGC-27 background, loss of DTX3L is expected to exacerbate genomic instability due to compromised DNA repair and to alter inflammatory signaling. This creates a powerful system for studying how ubiquitin ligases influence gastric cancer cell survival, immune evasion, and sensitivity to DNA-damaging agents. The model highlights the convergence of p53 deficiency and ubiquitin-mediated regulation, which is frequently dysregulated in gastrointestinal cancers.
Typical applications include functional dissection of DTX3L-dependent ubiquitin signaling, interferon responses, and NF-??B activation in gastric cancer cells; investigation of DNA damage repair pathways and genomic instability; drug sensitivity screening against DNA-damaging agents or immune modulators; and examination of tumor microenvironment interactions. Representative assays encompass Western blotting for DTX3L and phospho-STAT1, ubiquitination assays, co-immunoprecipitation of PARP9 complexes, and immunofluorescence for ??H2AX foci. Additional analyses include RNA-seq, migration/invasion assays, and apoptosis assays. For further information, please contact Ascent Research.
