The ADAMTS14 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma line. These cells harbor a targeted disruption of the ADAMTS14 gene, generated by transient expression of Cas9 and guide RNAs, resulting in a heterogeneous polyclonal pool with ADAMTS14 loss-of-function. As a polyclonal knockout, this product enables population-level analysis without clonal selection artifacts.
HT29 cells are a well-established intestinal epithelial cell model originally isolated from a primary colon adenocarcinoma resected from a 44-year-old female patient. This adherent cell line retains key features of colorectal adenocarcinoma and is widely utilized in cancer biology, gastrointestinal physiology, and drug transport studies due to its robust growth characteristics and compatibility with CRISPR-based genetic engineering.
ADAMTS14 encodes a secreted metalloprotease that functions as a procollagen N-proteinase, cleaving the N-terminal propeptides of procollagen types I, II, and III. This processing step is critical for collagen fibril assembly and extracellular matrix (ECM) remodeling. Transcription of ADAMTS14 is regulated by TGF-??, IL-1??, and the transcription factor SOX9, and is responsive to mechanical stress. Following activation, ADAMTS14 interacts with procollagen I, II, and III, along with collagen chaperones, to promote fibril formation. Downstream events involve lysyl oxidase-mediated collagen crosslinking and engagement of integrins, thereby connecting ADAMTS14 activity to cell adhesion and ECM integrity.
In the context of HT29 colorectal adenocarcinoma cells, disruption of ADAMTS14 impairs procollagen processing and collagen fibril formation, leading to aberrant ECM organization. This disturbance alters integrin-mediated adhesion and may compromise collective cell migration, a key process in tumor invasion. The polyclonal knockout pool models heterogeneous loss of ADAMTS14, mirroring the variable ECM remodeling observed in colorectal tumors.
These cells are suited for Western blotting and RT-qPCR to confirm knockout, collagen processing assays to measure enzymatic activity, and immunofluorescence to visualize ECM architecture. They support migration and invasion assays for metastasis research, as well as ECM adhesion studies. The model facilitates drug screening for ADAMTS14 or ECM-targeting compounds and aids functional genomics of the ADAMTS family in colorectal cancer. Contact Ascent Research for further information.
