ACTA1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for disruption of the ACTA1 gene in the HAP1 near-haploid human cell line. This product provides a robust loss-of-function model for investigating the cellular roles of alpha-skeletal muscle actin, encoded by ACTA1. The polyclonal format offers a heterogeneous knockout pool, enabling pooled screening and functional assays without clonal selection bottlenecks, making it suitable for studying diverse genetic perturbations within the same background.
The host cell line, HAP1, is derived from the KBM-7 chronic myeloid leukemia line and maintains a near-haploid karyotype, which facilitates efficient and unambiguous gene disruption in loss-of-function studies. HAP1 cells exhibit an adherent, fibroblast-like morphology and lack wild-type p53, contributing to their stable growth characteristics. These features have established HAP1 as a widely adopted platform for CRISPR-based screens, knockout validation, and pathway dissection in cancer biology and cell signaling research.
ACTA1 encodes the alpha-skeletal muscle actin isoform, a key structural component of sarcomeric thin filaments, where it interacts with tropomyosin, troponin, and nebulin to mediate muscle contraction. In non-muscle HAP1 cells, ACTA1 participates in broader actin cytoskeleton dynamics. The gene is transcriptionally regulated by serum response factor (SRF) and MEF2 transcription factors and functions within a network involving actin-binding proteins such as cofilin, profilin, vinculin, and tropomyosin 3, which modulate filament assembly, turnover, and linkage to adhesion complexes. Knockout of ACTA1 disrupts this network, leading to altered cytoskeletal organization, reduced cell?Cmatrix adhesion, and impaired motility, as supported by interactions with alpha-actinin and myosin heavy chains.
In the HAP1 cellular context, ACTA1 loss provides a non-muscle model that surrogates aspects of actin-related myopathies, including nemaline myopathy and actin-accumulation myopathy. The polyclonal knockout cells enable the study of ACTA1-dependent actin cytoskeletal defects without the need for muscle tissue, allowing high-throughput format screens. This model is particularly useful for dissecting the impact of ACTA1 deficiency on focal adhesion dynamics, cell migration, and cytoskeletal tension, as HAP1 cells rely on intact actin networks for their adherent phenotype.
Typical research applications encompass functional genomics screens, drug discovery studies targeting cytoskeletal pathways, and mechanistic investigations into actin-mediated processes. The polyclonal pool is amenable to assays such as immunofluorescence staining with phalloidin to visualize F-actin organization, Western blotting to assess ACTA1 and interacting protein levels, co-immunoprecipitation to probe binding partners, and cell migration assays including scratch wound and transwell formats. Additional applications include adhesion assays, proliferation measurements via MTT or cell counting, and FACS-based cell cycle analysis. For further information, please contact Ascent Research.
