The JADE1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line, offering a loss-of-function model for studying the epigenetic regulator JADE1. This product consists of a heterogeneous population of HT29 cells with targeted disruption of the JADE1 gene, generated using CRISPR/Cas9-mediated gene editing to introduce sequence alterations across the polyclonal pool. The polyclonal format preserves genetic diversity while achieving functional knockout at the population level, making it suitable for assays that do not require single-cell clonality. By ablating JADE1 expression, researchers can interrogate its role in chromatin modification, transcriptional regulation, and signaling networks in a well-characterized colorectal cancer background.
The HT29 host cell line is an established epithelial model of human colorectal adenocarcinoma, isolated from a primary tumor of a 44-year-old Caucasian female. HT29 cells exhibit a hyperdiploid karyotype with mutations in key oncogenes and tumor suppressors, including TP53 and APC, leading to constitutive activation of Wnt/??-catenin signaling. As an adherent, moderately differentiated cell line, HT29 is widely employed in cancer research to investigate proliferation, apoptosis, and drug responses. Its genetic profile and reproducible growth characteristics make it a reliable platform for functional genomics studies, particularly in the context of colorectal tumorigenesis.
JADE1 (Gene for Apoptosis and Differentiation in Epithelia) encodes a scaffold protein that serves as a core component of the HBO1 (KAT7) histone acetyltransferase complex, which also includes the adaptor proteins ING4 and ING5. Through this complex, JADE1 facilitates acetylation of histones H3 and H4, a post-translational modification tightly linked to transcriptional activation. JADE1 itself is regulated by upstream factors such as TP53, the VHL tumor suppressor, and hypoxia, while it exerts downstream effects on Wnt target genes including AXIN2, MYC, and CCND1. Mechanistically, JADE1 interacts directly with ??-catenin (CTNNB1) and is thought to bridge the HBO1 complex to Wnt-responsive promoters, thereby coordinating histone acetylation with TCF/LEF-mediated transcription. Additional pathway components include TCF7L2, a transcription factor that partners with ??-catenin to drive proliferative gene programs. Disruption of JADE1 is predicted to uncouple histone acetylation from Wnt signaling, altering the expression of genes governing cell cycle progression and survival.
In the HT29 colorectal adenocarcinoma background, JADE1 knockout provides a contextually relevant system to dissect the intersection of epigenetic regulation and oncogenic Wnt signaling. Given that HT29 cells harbor APC mutations leading to ??-catenin stabilization, loss of JADE1 may further modulate the output of this pathway, potentially affecting downstream targets such as MYC and CCND1 that are critical for tumor cell proliferation. This model thus enables the investigation of JADE1 as a potential tumor suppressor or oncogenic cofactor in colon cancer, and allows for the assessment of how alterations in histone acetylation patterns impact colorectal cancer cell behavior. The interplay between VHL, hypoxia, and JADE1 can also be explored, as HT29 cells are responsive to hypoxic conditions, offering insights into microenvironmental regulation of epigenetic complexes.
This knockout tool is suited for a broad range of experimental designs. Applications include profiling JADE1-dependent transcriptional changes via RNA-seq, quantifying Wnt pathway activity using TOP/FOP flash reporter assays, and mapping JADE1 chromatin occupancy through ChIP-qPCR. Functional assays such as colony formation, migration, and flow cytometry-based apoptosis assays can delineate JADE1??s influence on cell proliferation and death. Co-immunoprecipitation and western blotting enable validation of protein interactions with HBO1, ING4/ING5, and ??-catenin. Additionally, the cells can serve in epigenetic drug screening to identify compounds that modulate acetyltransferase complex function or phenocopy JADE1 loss. For further information and technical support, please contact Ascent Research.
