The BAIAP2L1 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population for studying BAIAP2L1 gene function. It consists of a heterogeneous pool of HAP1 cells that have undergone CRISPR/Cas9-mediated disruption of the BAIAP2L1 locus, leading to functional inactivation of the encoded IRTKS protein. As a polyclonal population, the cells reflect diverse editing outcomes while enabling robust loss-of-function experiments without requiring clone isolation. This format supports both pooled screening applications and bulk cell-based assays, offering flexibility for functional genomics studies.
The HAP1 cell line is a near-haploid human cell line derived from the KBM-7 chronic myelogenous leukemia line. It retains a single copy of most chromosomes, except for a heterozygous 30 Mb fragment of chromosome 15, making it an ideal host for haploid genetic screens and gene disruption studies. HAP1 cells are widely utilized in drug testing, knockout-based functional analysis, and high-throughput screening due to their ease of genetic manipulation and the straightforward genotype?Cphenotype correlation afforded by their hemizygous background. This genetic simplicity accelerates the generation of interpretable loss-of-function phenotypes.
BAIAP2L1 encodes insulin receptor tyrosine kinase substrate (IRTKS), a multifunctional scaffold protein that bridges small GTPases to the actin cytoskeleton. IRTKS directly interacts with CDC42 and RAC1 and functions downstream of the insulin receptor and growth factor signals such as IGF-1. Mechanistically, IRTKS links activated GTPases to the WAVE2?CARP2/3 complex, driving actin polymerization and formation of filopodia and lamellipodia. It also acts as a substrate of the insulin receptor, integrating metabolic signals with cytoskeletal reorganization. Key interacting partners include BAI1, IRSp53 (BAIAP2), EPS8, WAVE2, and F-actin, while downstream effectors encompass the ARP2/3 complex and PI3K?CAKT signaling.
Disruption of BAIAP2L1 in the HAP1 near-haploid background creates a valuable model to dissect IRTKS-dependent pathways in a leukemic context. Since HAP1 cells express a single copy of BAIAP2L1, the polyclonal knockout population likely yields loss-of-function phenotypes that are free from compensatory contribution by a second allele, enhancing the clarity of functional readouts. This system facilitates exploration of how IRTKS coordinates migratory and invasive behavior in cancer, including its potential roles in hematopoietic malignancies and solid tumor metastasis, bridging cell biology with translational cancer research.
This knockout model is suited for a wide range of experimental approaches. Researchers can employ Transwell migration and invasion assays to quantify BAIAP2L1-dependent motility, complemented by live-cell imaging to visualize filopodia dynamics. Western blotting and RT-qPCR confirm IRTKS disruption, while co-immunoprecipitation with CDC42, RAC1, or BAI1 validates interaction networks. Immunofluorescence using phalloidin staining can reveal F-actin structural changes, and phospho-AKT analysis assesses insulin signaling output. These applications position the BAIAP2L1 Knockout HAP1 Polyclonal Cells as a robust tool for functional studies in cancer biology, cell migration, and insulin receptor signaling. For further information, please contact Ascent Research.
