HMOX1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma line. This product introduces a loss-of-function model of the HMOX1 gene through targeted gene disruption, generating a heterogeneous pool of edited cells that retains population diversity. It is designed for functional genomics, pathway analysis, and drug screening in a cancer-relevant epithelial context, providing a robust tool for investigating HO-1-dependent cytoprotective mechanisms.
The HT29 cell line, isolated from a primary colorectal adenocarcinoma, is an extensively used epithelial model with mutant p53 and responsiveness to TGF-??. These cells exhibit an adherent, undifferentiated morphology that can be induced to differentiate, and they are endowed with active NRF2 signaling. HT29 is a standard host for studying colorectal cancer biology, oxidative stress responses, and inflammatory signaling, making it ideal for examining HO-1-mediated processes.
HMOX1 encodes heme oxygenase-1 (HO-1), the rate-limiting enzyme in heme catabolism. Under the control of the KEAP1-NRF2-ARE pathway and transcriptional repressor Bach1, HO-1 is induced by stimuli such as heme, cobalt protoporphyrin, IL-10, and TGF-??. HO-1 cleaves pro-oxidant heme into biliverdin, carbon monoxide (CO), and free iron. Biliverdin is reduced to the potent antioxidant bilirubin by biliverdin reductase; CO activates soluble guanylyl cyclase to produce cGMP and modulates MAPK and PI3K/AKT cascades; iron is sequestered by ferritin. HO-1 additionally interacts with Caveolin-1 and negatively regulates NF-??B, collectively exerting anti-inflammatory, antiapoptotic, and cytoprotective effects.
In HT29 colorectal cancer cells, HO-1 is implicated in chemoresistance, redox homeostasis, and ferroptosis inhibition. HMOX1 disruption permits dissection of how biliverdin/bilirubin, CO, and iron handling influence tumor cell survival, lipid peroxidation, and inflammatory crosstalk. The mutant p53 and TGF-??-responsive background of HT29 further enables study of interactions between HO-1 and DNA damage responses or epithelial-mesenchymal transition. This model is thus valuable for identifying HO-1-dependent oncogenic mechanisms and potential therapeutic targets in colorectal carcinoma.
Representative applications include Western blotting and RT-qPCR for HO-1 expression validation, ROS measurement, and ferroptosis lipid peroxidation assays using erastin or RSL3. Cell viability assays following hemin treatment, wound healing migration studies, and drug sensitivity profiling with NRF2 activators (e.g., sulforaphane) or HO-1 inhibitors (e.g., tin protoporphyrin) are facilitated. CO signaling can be probed via cGMP quantification, and anti-inflammatory effects assessed with IL-10. For further information, please contact Ascent Research.
