The JAM3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population engineered from the HT29 human colorectal adenocarcinoma cell line, featuring targeted disruption of the JAM3 gene. This loss-of-function model enables researchers to interrogate the biological roles of JAM3 in epithelial biology, barrier regulation, and disease pathogenesis without the confounding effects of residual protein expression. As polyclonal cells, the population comprises a heterogeneous pool of edited alleles, offering a robust system for studying gene function at the population level.
The parental HT29 cell line originates from a primary tumor of a 44-year-old Caucasian female with colorectal adenocarcinoma. HT29 cells exhibit epithelial morphology and are capable of differentiation into enterocyte- and mucus-secreting phenotypes under appropriate culture conditions. This cell line is widely employed in cancer research, intestinal barrier studies, and investigations of drug absorption, making it an ideal host for modeling JAM3-related phenotypes in a colorectal cancer setting.
JAM3 encodes a junctional adhesion molecule that localizes to tight junctions and mediates cell-cell adhesion, leukocyte transmigration, and angiogenesis. It interacts with JAM2 and integrin ??M??2 (Mac-1) to regulate paracellular permeability and immune cell trafficking. Intracellularly, JAM3 associates with PDZ-domain scaffold proteins ZO-1, afadin, PAR-3, and PAR-6, coupling to actin dynamics via Cdc42 and Rac1 activation. Upstream regulators include VEGF/VEGFR2 signaling, inflammatory cytokines (TNF-??, IL-1??), mechanical forces, and the Sp1 transcription factor.
In the HT29 colorectal adenocarcinoma context, JAM3 knockout disrupts tight junction integrity and may enhance paracellular permeability, providing a valuable model for studying epithelial barrier dysfunction and tumor cell invasion. Given HT29??s ability to form polarized monolayers with measurable transepithelial electrical resistance (TEER), this knockout system is particularly suited for dissecting the contribution of JAM3 to colorectal cancer progression, metastasis, and the inflammatory tumor microenvironment. Additionally, the model can be applied to investigate the molecular underpinnings of JAM3-associated syndromes like hemorrhagic destruction of the brain, subependymal calcification, and cataracts.
Typical applications include TEER and FITC-dextran permeability assays for barrier function, Boyden chamber migration/invasion assays, and cell adhesion assays. Western blotting and immunofluorescence confirm JAM3 loss and localize junctional proteins. Flow cytometry and co-immunoprecipitation assess interacting partners like ZO-1 and integrin ??M??2. The model is also suitable for reovirus binding assays. For further information, please contact Ascent Research.
