The AIF1L Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-mediated gene disruption model designed to dissect the biological functions of the AIF1L gene in cytoskeletal regulation and cell migration. Supplied as a polyclonal knockout cell pool, this product contains a heterogeneous mix of HAP1 cells harboring diverse loss-of-function edits at the target locus, offering a convenient, high-throughput-compatible resource for phenotypic screening without the need for single-cell cloning.
The parental HAP1 line is a human near-haploid chronic myeloid leukemia-derived cell model with adherent fibroblastoid morphology. Its near-haploid genomic architecture simplifies knockout generation and reduces functional redundancy, enabling clearer attribution of phenotypes to gene disruption. HAP1 is widely employed in hematopoietic research, cancer biology studies, and genome-scale screening initiatives due to its stable karyotype and retention of key myeloid lineage signaling networks.
AIF1L encodes a putative actin-bundling protein that interacts with F-actin and is responsive to intracellular Ca2+ and Rho GTPase signals. The gene is transcriptionally activated by pro-inflammatory mediators including TNF-??, IL-1??, and TGF-??, primarily through NF-??B-dependent mechanisms. AIF1L protein functions downstream of these stimuli to promote actin polymerization, turnover of focal adhesion complexes, and enhanced cell motility, while also modulating matrix metalloproteinase (MMP) expression. It operates within a signaling cascade involving RhoA, ROCK, LIMK, and Cofilin to orchestrate actin filament stabilization and dynamic reorganization.
Knockout of AIF1L in the HAP1 background disrupts the linkage between inflammatory signaling and the cytoskeletal machinery, providing a powerful tool for studying myeloid cell invasiveness and the tumor microenvironment. The polyclonal pool mimics a naturally heterogeneous knockout population, allowing researchers to evaluate the spectrum of migration, adhesion, and morphological defects that arise from varied mutation types. This feature is particularly valuable for modeling cancer metastasis, where diverse clones within a tumor exhibit differential invasive capabilities.
Researchers can use these cells in a variety of assays, including Western blotting to confirm AIF1L depletion and downstream phospho-signaling (e.g., phospho-Cofilin), immunofluorescence visualization of F-actin stress fibers, and quantitative transwell migration/invasion assays. Additional applications encompass RNA-sequencing for transcriptomic profiling, adhesion assays on extracellular matrix substrates, and flow cytometry for surface integrin or cytokine receptor expression. The model is well-suited for drug target validation in anti-metastatic and anti-inflammatory therapeutic research. For further technical details or custom requests, please contact Ascent Research.
