CBLL1 Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colon adenocarcinoma epithelial line. This pool enables loss-of-function studies of CBLL1, encoding the E3 ubiquitin ligase Hakai, by disrupting endogenous gene expression. The model provides a robust system to investigate CBLL1??s roles in cell adhesion, epithelial-mesenchymal transition (EMT), and cancer progression. As a non-clonal population, it captures editing heterogeneity, making it suitable for studies requiring averaged phenotypes or avoidance of clonal artifacts.
HT29 cells, isolated from a primary colon adenocarcinoma of a 44-year-old female, are a standard model for colorectal cancer and intestinal epithelial biology. They form polarized monolayers with tight and adherens junctions, expressing high E-cadherin levels, and are thus ideal for examining barrier function, cell adhesion dynamics, EMT, and metastasis. The epithelial context and well-characterized signaling networks make HT29 particularly relevant for dissecting CBLL1 function in colorectal cancer.
CBLL1 (Hakai) is an E3 ubiquitin ligase that ubiquitinates E-cadherin, targeting it for proteasomal degradation and thereby weakening adherens junctions to promote EMT. Its activity is stimulated by upstream regulators including Src kinase, growth factors (EGF, HGF), and TGF-beta. CBLL1 interacts with E-cadherin, p120-catenin, and Cbl proteins to execute ubiquitination. E-cadherin loss releases beta-catenin, which can enter the nucleus and activate Wnt-responsive EMT genes. Thus, CBLL1 coordinates adhesion dynamics with transcriptional reprogramming during invasion.
In HT29 cells, which display robust epithelial features, CBLL1 knockout likely stabilizes E-cadherin and reinforces cell-cell contacts, potentially blocking EMT initiation. This loss-of-function model allows interrogation of CBLL1 in colorectal cancer progression, metastasis, and anoikis resistance. Experiments comparing knockout and wild-type cells can delineate Hakai-mediated junctional turnover and link Src and Wnt/beta-catenin signals to adhesive changes in a colon cancer milieu.
This polyclonal knockout population supports diverse assays: Western blot and flow cytometry for E-cadherin quantification, immunofluorescence for junctional integrity, scratch wound and transwell invasion assays for cell motility, and RT-qPCR for EMT markers (e.g., SNAIL, VIMENTIN). Ubiquitination assays can directly probe E-cadherin modification. Researchers in colorectal cancer biology, EMT, metastasis, and ubiquitin-proteasome systems will find this model useful for mechanistic and therapeutic studies. For further information, please contact Ascent Research.
