The ADAMTS14 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the ADAMTS14 gene in the HCT 116 human colorectal carcinoma cell line. This loss-of-function model enables researchers to study the role of ADAMTS14 in extracellular matrix (ECM) remodeling and colorectal cancer biology without relying on pharmacological inhibition. The polyclonal format provides a heterogeneous knockout pool that maintains genetic diversity while collectively abolishing ADAMTS14 protein expression, making it suitable for population-level functional studies.
The host HCT 116 cell line is an adherent epithelial cell line derived from a 58-year-old male with colorectal carcinoma. It exhibits microsatellite instability-high (MSI-H) status and harbors activating mutations in KRAS (G13D), PIK3CA, and CTNNB1, which are common in colorectal cancer. These genetic alterations drive constitutive activation of the RAS?CMAPK, PI3K?CAKT, and WNT signaling pathways, establishing HCT 116 as a robust model system for studying oncogenic signaling and tumor progression. Its well-characterized behavior in adhesion, migration, and invasion assays further enhances its utility.
ADAMTS14 encodes a procollagen N-proteinase critical for collagen maturation and ECM remodeling. It processes procollagens type I and II by cleaving their N-propeptides, facilitating proper fibril assembly. ADAMTS14 expression and activity are regulated by upstream factors including TGF-beta, SMAD2/3, IL-1beta, TNF-alpha, and WNT3A. Downstream, it promotes the deposition of collagen type I and II and aggrecan. Its function is modulated through interactions with tissue inhibitor of metalloproteinases 3 (TIMP-3), collagen type I, fibronectin, and aggrecan. Representative pathway components linking ADAMTS14 to cell-matrix signaling include COL1A1, the integrin heterodimer ITGA2/ITGB1, TGF-beta receptor I (TGFBR1), SMAD2, focal adhesion kinase (FAK), and SRC kinase.
In the context of HCT 116 cells, ADAMTS14 knockout provides a platform to dissect the interplay between ECM remodeling and oncogenic signaling pathways such as TGF-beta, PI3K-AKT, and integrin-mediated adhesion. Given the host line??s mutant KRAS and PI3K, loss of ADAMTS14 may perturb collagen processing, alter cell?Cmatrix interactions, and modulate migratory and invasive properties. This model is particularly relevant for examining how ECM proteolysis influences colorectal cancer progression, including metastasis, where ECM integrity and signaling are often dysregulated.
Research applications include characterizing ADAMTS14??s role in colorectal cancer ECM remodeling using western blotting for collagen processing and immunofluorescence for ECM components. Functional studies of cell migration and invasion can be performed with wound healing and Transwell invasion assays, while adhesion assays on collagen allow assessment of integrin-dependent attachment. TGF-beta signaling reporter assays and RNA-seq transcriptomic analysis enable elucidation of downstream signaling changes. This model supports drug target validation for anti-metastatic therapies and functional genomics in cancer biology. For further information, please contact Ascent Research.
