HM13 Knouckout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human colorectal adenocarcinoma HT29 cell line. This product provides a heterogeneous pool of HM13-disrupted cells, enabling researchers to study loss-of-function effects across a diverse genetic background. The polyclonal nature avoids clonal bias and better captures population-level responses, making it suitable for functional genomics and pathway analysis in intestinal epithelial biology and colorectal cancer research. The CRISPR/Cas9-mediated gene disruption generates a robust model to interrogate the roles of intramembrane proteolysis in cancer and immune signaling without claiming complete or biallelic knockout, ensuring applicability for pooled screening and validation workflows.
The HT29 cell line is a well-established model of human colorectal adenocarcinoma with epithelial morphology, extensively utilized to investigate intestinal cell differentiation, tumorigenesis, and mucosal immunology. Isolated from a primary tumor of a 44-year-old female, HT29 cells can undergo enterocytic differentiation under specific culture conditions, providing a versatile platform for studying colorectal cancer progression and epithelial barrier function. Their genetic background, including mutations in APC and TP53, renders them particularly relevant for dissecting oncogenic signaling and inflammatory crosstalk in the intestinal epithelium, making this knockout derivative a powerful tool for target validation and mechanistic studies.
HM13 (SPPL2A) encodes an intramembrane protease that cleaves type II transmembrane proteins, primarily TNF?? and the invariant chain CD74, thereby modulating their downstream signaling outputs. The enzyme is activated by cellular stress signals and regulated by upstream factors such as TNF?? and IL-1??. Upon cleavage, it releases soluble TNF?? fragments and the CD74 intracellular domain, which can influence NF-??B transcriptional activity and Notch1 signaling through interactions with presenilin (PSEN1) and signal peptide peptidase. HM13-mediated proteolysis sits at the intersection of the TNF?? signaling, CD74/MIF signaling, and intramembrane proteolysis pathways, controlling antigen processing and presentation as well as inflammatory responses. Key pathway components include TNF??, CD74, MIF, NF-??B, Notch1, and PSEN1, collectively shaping immune surveillance and epithelial homeostasis.
In the HT29 colorectal cancer context, HM13 disruption impairs intramembrane proteolysis of substrates like CD74, leading to altered antigen presentation and potentially dysregulated NF-??B and Notch signaling. This knockout model is particularly significant for investigating mechanisms of inflammatory bowel disease?Cassociated cancer progression, where HM13-dependent processing of immune modulators influences tumor microenvironments. The interaction between HM13 and CD74 is critical for regulating surface expression of MHC class II complexes and cell fate decisions; thus, this model enables dissection of how epithelial cells coordinate innate immune signals and antigen handling. Furthermore, by removing HM13 activity, researchers can explore the crosstalk between proteolytic cleavage events and oncogenic pathways, offering insights into therapeutic vulnerabilities in colorectal cancer and immunodeficiency syndromes.
Typical research applications encompass functional studies of intramembrane proteolysis in inflammation-associated carcinogenesis, validation of HM13 as a drug target for immune modulation, and dissection of antigen presentation pathways in intestinal epithelial cells. Representative assays include Western blotting to detect HM13 and substrate cleavage products, RT-qPCR for expression profiling, ELISA to quantify TNF?? secretion, flow cytometry for CD74 surface expression, NF-??B luciferase reporter assays to monitor transcriptional activity, and migration/invasion assays to assess metastatic potential. Apoptosis assays can further clarify the role of HM13 in cell survival. This polyclonal knockout population is suitable for both arrayed and pooled screening formats, facilitating high-throughput discovery pipelines. For further information, to inquire about custom specifications, or to obtain technical support, please contact Ascent Research.
