The BASP1 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-mediated polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, engineered for loss-of-function investigation of the BASP1 gene. This polyclonal population contains a heterogeneous mixture of edited alleles, enabling studies of gene disruption effects without clonal selection bias and is supplied as a frozen vial of early-passage cells.
HT29 is an epithelial colorectal adenocarcinoma cell line derived from a primary colon tumor of a 44-year-old female patient. It harbors well-characterized mutations in APC, BRAF (V600E), and PIK3CA, which drive constitutive activation of MAPK/ERK and PI3K/AKT/mTOR oncogenic pathways. The cells exhibit chromosomal instability and are tumorigenic in immunocompromised mice, establishing HT29 as a robust model for studying colorectal cancer signal transduction, targeted therapy response, and metastatic progression.
BASP1 is a membrane-associated protein involved in actin cytoskeleton regulation, cell motility, and transcriptional co-regulation. It binds calmodulin and sequesters phosphatidylinositol 4,5-bisphosphate (PIP2), linking membrane phospholipid metabolism to actin filament dynamics. BASP1 activity is regulated by upstream kinases including Protein Kinase C (PKC) and SRC kinase, and it serves as a co-regulator for the transcription factors WT1 and EBF1. Interactions with N-myristoyltransferase and actin-binding proteins further position BASP1 at the interface between signaling and structural reorganization. Downstream, BASP1 influences expression of GAP-43 and WT1/EBF1 target genes such as MYC and CCND1. In the HT29 context, where BRAF V600E and PIK3CA mutations hyperactivate the MAPK/ERK and PI3K/AKT/mTOR cascades, BASP1 knockout is predicted to disrupt PIP2 sequestration and calmodulin-mediated processes, thereby affecting actin remodeling, ERK1/2, AKT, mTOR, ??-catenin, and cofilin activity.
The combination of BRAF V600E-driven MAPK signaling and PIK3CA-mediated PI3K activation makes HT29 cells highly dependent on cytoskeletal control for migration and invasion. BASP1 loss in this genetic background provides a powerful model to dissect how membrane-proximal actin regulation integrates with oncogenic transcription programs to modulate colorectal cancer cell behavior, EMT, and metastatic potential. This system can help unravel context-specific dependencies that may be exploited for therapeutic intervention.
These polyclonal knockout cells are ideally suited for functional assays such as Transwell migration/invasion studies, immunofluorescence imaging of actin stress fibers, and proliferation measurements via MTT or CCK-8. They enable investigation of drug sensitivity, particularly to BRAF inhibitors (e.g., vemurafenib) and PI3K/mTOR inhibitors, and can be used in combination with RNA-seq to map BASP1-dependent transcriptomic changes. Additional applications include flow cytometric cell cycle analysis and neurobiology-directed differentiation protocols to study neurite outgrowth. For further technical details, please contact Ascent Research.
