The ITGA1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the human ITGA1 gene. This heterogeneous pool of HT29 cells has undergone CRISPR/Cas9-mediated disruption of the ITGA1 genomic locus, abolishing integrin alpha-1 subunit expression. The polyclonal format provides a population-level knockout model, enabling collective assessment of cellular responses to ITGA1 inactivation without clonal selection. This product serves as a robust tool for investigating integrin-mediated processes in a colorectal adenocarcinoma context.
The HT29 host cell line is a well-characterized human colorectal adenocarcinoma model derived from a 44-year-old Caucasian female. These adherent cells display epithelial morphology and are distinguished by their mucin-producing phenotype, making them a valuable intestinal epithelial model. HT29 cells are widely employed in cancer research to study tumorigenesis, differentiation, and therapeutic responses. Their consistent molecular profile and reproducible growth provide a reliable background for gene editing, facilitating direct comparisons between wild-type and ITGA1-knockout populations.
ITGA1 encodes the integrin alpha-1 subunit, which heterodimerizes with integrin beta-1 (ITGB1) to form a receptor for collagen and laminin. Upon ligand binding, the complex recruits talin, kindlin, and paxillin, triggering autophosphorylation of focal adhesion kinase (FAK) and activation of Src family kinases. This FAK/Src axis phosphorylates p130Cas and Crk, leading to downstream activation of the ERK and PI3K-Akt pathways, with mTOR as a key effector. ITGA1 expression is transcriptionally regulated by TGF-beta, IL-1, TNF-alpha, SP1, and AP-1, while its signaling modulates MMP9 and cyclin D1, thereby controlling cell adhesion, migration, proliferation, and survival.
In HT29 colorectal adenocarcinoma cells, ITGA1 knockout disrupts integrin-dependent interactions with collagen IV and laminin, critical components of the tumor microenvironment. Loss of alpha-1 integrin function impairs cell adhesion, spreading, and migration, providing a reductionist model to dissect ITGA1??s roles in tumor invasion, metastasis, and stromal crosstalk. Given HT29??s mucin-producing nature, this model also offers insight into epithelial barrier dysfunction and inflammatory signaling, relevant to fibrosis, rheumatoid arthritis, and inflammatory bowel disease.
Researchers can employ this polyclonal knockout population in diverse assays, including Western blotting and RT-qPCR for knockout confirmation, flow cytometry for surface integrin profiling, and adhesion assays on collagen I/IV or laminin. Migration and invasion are assessed via Boyden chamber assays, proliferation via MTT, and focal adhesion dynamics by immunofluorescence for phospho-FAK and paxillin. This model is particularly suited for anti-metastatic drug screening, ECM-receptor interaction studies, and mechanistic investigations of ITGA1-dependent pathways in colorectal cancer. For inquiries or custom requirements, contact Ascent Research.
