The EFCAB14 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells that carry targeted disruptions of the EFCAB14 gene. This product provides a loss-of-function model for EFCAB14, a gene encoding an EF-hand calcium-binding domain-containing protein, without the clonal isolation step, thereby preserving the heterogeneity of editing outcomes. The polyclonal format is well-suited for initial functional screening, pooled assays, and experiments where the average effect of gene disruption is assessed across a genetically diverse cell population.
The host cell line, HAP1, is a near-haploid human cell line derived from the chronic myeloid leukemia cell line KBM-7. Its near-haploid karyotype simplifies recessive genetic screens and CRISPR-based knockout experiments by minimizing genetic redundancy. Originating from a male donor, HAP1 cells retain key signaling machinery of hematopoietic lineages and are widely employed for studying gene function in a simplified genomic context, including calcium signaling and cytoskeletal dynamics.
EFCAB14 is classified as an EF-hand calcium-binding protein, implying a role in sensing intracellular calcium fluctuations and translating them into downstream cellular responses. Within the calcium signaling network, EFCAB14 is predicted to be regulated by intracellular calcium levels and interact with calmodulin, tropomyosin, and myosin light chain kinase. It functions downstream of calmodulin and CaMKII, and its signaling converges on effectors such as ??-actin (ACTB), myosin heavy chain 9 (MYH9), calpain, and calcineurin. These molecular associations position EFCAB14 at a node connecting calcium homeostasis to the regulation of the actin cytoskeleton, with potential influence on cell shape, adhesion, and contractility.
In the HAP1 context, disruption of EFCAB14 may unmask phenotypes related to calcium-dependent cytoskeletal reorganization, cell motility, and survival, as the near-haploid state reduces compensatory gene expression. By eliminating EFCAB14 function, researchers can interrogate its contribution to calcium-mediated processes such as migration, adhesion turnover, and apoptosis, all of which are relevant to cancer cell biology and hematological malignancies. The interaction network involving calmodulin, tropomyosin, and myosin regulatory pathways further suggests that EFCAB14 loss may alter actomyosin dynamics and cellular mechanics.
This knockout model is amenable to a broad spectrum of experimental approaches, including Western blotting and RT-qPCR for confirming target disruption, immunofluorescence and live-cell imaging for visualizing cytoskeletal changes, calcium flux assays to assess signaling competence, and migration or wound-healing assays to evaluate cell motility. Additionally, cell cycle analysis and apoptosis assays can reveal roles in proliferation and survival. The EFCAB14 Knockout HAP1 Polyclonal Cells thus serve as a versatile tool for functional genomics, drug discovery screens, and mechanistic studies of calcium signaling in a near-haploid background. For more information, please contact Ascent Research.
