The BAIAP2L1 Knockout HT29 Polyclonal Cells are a heterogeneous population of HT29 colorectal adenocarcinoma epithelial cells with CRISPR/Cas9-mediated disruption of the BAIAP2L1 gene. This polyclonal knockout model provides a pooled loss-of-function system that captures genetically diverse edits, circumventing clonal selection bias; the cells maintain characteristic HT29 adherent growth.
HT29 cells originate from a human colorectal adenocarcinoma of a female donor and are a standard intestinal epithelial model. These adherent, differentiation-competent cells are extensively used in colorectal cancer research to investigate tumorigenesis, metastasis, and epithelial barrier function. The disease-relevant background is apt for interrogating BAIAP2L1, which is overexpressed in several gastrointestinal cancers and linked to aggressive phenotypes.
BAIAP2L1 is an adaptor protein that couples receptor tyrosine kinase signals??from insulin receptor (IR), IGF-1 receptor, and EGFR??to actin cytoskeleton remodeling. It acts downstream of IR/IRS and Src family kinases, interacting with IRS-1, Src, WAVE2, and Abi1 to promote Arp2/3-mediated actin polymerization. The IR??IRS??BAIAP2L1??WAVE2/Abi1??Arp2/3 cascade and integrin??Src??BAIAP2L1??Rac1??WAVE pathway collectively regulate membrane protrusion, cell migration, and invasion, contributing to cancer progression.
In HT29 cells, BAIAP2L1-dependent actin dynamics are critical for migration and metastatic potential. Disrupting BAIAP2L1 in this polyclonal population enables dissection of its specific roles in insulin-stimulated motility, cytoskeletal reorganization, and tumor progression, without clonal artifacts. The model retains xenograft tumorigenicity, facilitating translation to in vivo metastasis studies.
This knockout model supports a range of assays: wound healing and Transwell migration/invasion to quantify motility, F-actin immunofluorescence to visualize cytoskeletal changes, and co-immunoprecipitation with IRS-1 or Src to probe interactions. Insulin-stimulated phospho-Akt detection and MTT/BrdU proliferation assays evaluate signaling and growth outputs. The cells are suitable for inhibitor screening targeting BAIAP2L1 pathways and epithelial barrier assessments using TEER. For further details, please contact Ascent Research.
