ALDH1A3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma epithelial cell line, designed to disrupt the ALDH1A3 gene. This product comprises a heterogeneous pool of edited cells, enabling robust loss-of-function studies of ALDH1A3 without the need for single-cell cloning. The polyclonal format retains population-level diversity, providing a practical and physiologically relevant model for investigating retinoid signaling in colorectal cancer research.
The HT29 cell line, originally isolated from a primary colorectal adenocarcinoma of a 44-year-old female, harbors a mutant p53 protein and serves as a foundational in vitro model for colorectal carcinoma. These cells display an epithelial morphology and are capable of enterocytic differentiation under appropriate culture conditions, making them valuable for studying intestinal epithelial barrier function, tumorigenesis, and chemotherapeutic response. HT29 cells are extensively characterized and widely employed in cancer biology for drug screening and mechanistic pathway analysis.
ALDH1A3 encodes an aldehyde dehydrogenase that irreversibly oxidizes retinaldehyde to all-trans retinoic acid, the principal active retinoid that binds nuclear retinoic acid receptors RAR/RXR. These ligand-activated transcription factors regulate a broad gene network, including HOX clusters and p21, and are critical for stem cell maintenance, differentiation, and epithelial-mesenchymal transition. ALDH1A3 expression is transcriptionally regulated by upstream signals such as Wnt/??-catenin, NF-??B, STAT3, and hypoxia-inducible factors. The synthesized retinoic acid interacts with cellular retinoic acid binding proteins (CRABP1/2) for intracellular trafficking and is catabolized by CYP26 enzymes, forming a tightly controlled signaling node. Downstream, RAR/RXR activation sustains stemness markers like Sox2, Oct4, and Nanog, linking ALDH1A3 activity directly to cancer stem cell phenotypes.
In HT29 colorectal cancer cells, ALDH1A3 contributes to the maintenance of a chemoresistant stem cell pool by fueling retinoic acid biosynthesis. Disruption of ALDH1A3 in this polyclonal knockout model attenuates RAR/RXR-dependent transcriptional programs, leading to diminished expression of stemness and anti-apoptotic factors. This knockout thus abrogates a key pathway supporting undifferentiated tumor cell survival and drug efflux mechanisms. The HT29 background provides a relevant context to study how ALDH1A3-mediated retinoid signaling intersects with colorectal cancer progression, differentiation, and therapeutic resistance.
This knockout product is ideally suited for diverse investigations including colorectal cancer stem cell biology, retinoic acid signaling dynamics, and chemoresistance mechanisms. Researchers can employ ALDEFLUOR flow cytometry to assess ALDH enzymatic activity, LC-MS for retinoic acid quantification, sphere formation assays to evaluate stemness, and chemosensitivity panels using MTT or drug efflux assays. Transcriptional profiling via RNA-seq and targeted RT-qPCR for RAR/RXR downstream targets further elucidate pathway alterations. These tools facilitate differentiation therapy development and cancer metabolism studies. For technical inquiries or ordering information, please contact Ascent Research.
