The ALDH1B1 Knockout HT29 Polyclonal Cells product provides a heterogeneous population of HT29 human colorectal adenocarcinoma cells engineered with CRISPR/Cas9-mediated disruption of the Aldehyde Dehydrogenase 1 Family Member B1 (ALDH1B1) gene, generating a loss-of-function model for studying mitochondrial aldehyde dehydrogenase activities. This polyclonal knockout format preserves genetic diversity within the edited pool, avoiding biases introduced by single-cell cloning while effectively reducing ALDH1B1 expression across the population. The targeted gene disruption is achieved without introducing exogenous sequences, maintaining the native genomic context for downstream analyses.
The host HT29 cell line originates from a human colorectal adenocarcinoma of a female donor and is well-characterized as a model for intestinal epithelial biology. These cells exhibit properties of absorptive and mucus-secreting enterocytes, capable of forming polarized monolayers and producing mucins such as MUC2 under differentiation conditions. HT29 cells are extensively used to study epithelial differentiation, barrier function, and the cellular responses to xenobiotics and metabolites, making them a relevant system for addressing colorectal cancer biology and gut physiology.
ALDH1B1 encodes a mitochondrial aldehyde dehydrogenase that catalyzes the NAD+-dependent oxidation of a range of aldehyde substrates, notably retinaldehyde and acetaldehyde. The conversion of retinaldehyde to retinoic acid is a critical source of this morphogen, which activates nuclear receptors RAR/RXR to transcriptionally regulate genes like CYP26A1 and HOX cluster members, thereby controlling cellular differentiation and proliferation. ALDH1B1 expression is itself regulated by upstream factors including HNF4A, PPAR??, RAR/RXR, and NFE2L2, and it interacts with cofactor NAD+ and substrates retinaldehyde and acetaldehyde. Additionally, ALDH1B1 participates in ethanol degradation by detoxifying acetaldehyde, linking its function to alcohol metabolism and protection against reactive aldehydes.
Disruption of ALDH1B1 in HT29 cells is anticipated to impair retinoic acid synthesis, altering the signaling cascade that governs intestinal epithelial differentiation. This defect can affect mucin production, tight junction integrity, and the expression of differentiation markers such as MUC2, while also compromising the cell’s ability to neutralize cytotoxic aldehydes like acetaldehyde. The knockout model therefore offers a platform to examine how aberrant ALDH1B1-dependent metabolism contributes to colorectal adenocarcinoma maintenance, response to oxidative stress, and the pathophysiological intersection of alcohol consumption and colon cancer.
This polyclonal knockout cell population is suitable for a broad range of functional and molecular assays, including Western blotting and RT-qPCR for ALDH1B1 and retinoic acid-responsive genes, HPLC-based retinoic acid quantification, aldehyde dehydrogenase activity measurements, MTT proliferation assays, immunofluorescence for MUC2 localization, cell migration studies, and transcriptomic profiling via RNA-seq. Researchers can apply these tools to investigate retinoic acid signaling in colorectal cancer, the role of aldehyde detoxification in intestinal epithelium, and the crosstalk between ethanol metabolism and colon carcinogenesis. For further details or technical support, please contact Ascent Research.
