The EFHD1 Knockout HAP1 Polyclonal Cells are a polyclonal CRISPR/Cas9-edited HAP1 cell population with targeted disruption of the EFHD1 gene, creating a loss-of-function model for studying EFHD1’s role in adhesion and migration. This polyclonal pool avoids clonal selection artifacts and enables robust functional studies. The CRISPR/Cas9-mediated gene disruption eliminates functional EFHD1 protein, providing a versatile tool for signal transduction research.
HAP1 cells are a near-haploid human cell line derived from chronic myelogenous leukemia KBM-7 cells, widely used for haploid genetic screening due to their single-allele status. They exhibit adherent, fibroblast-like morphology and support standard cell culture and diverse downstream assays. The haploid karyotype minimizes genetic redundancy, facilitating clean knockout models and unambiguous phenotypic readouts.
EFHD1 is a calcium-binding scaffold protein that bridges calcium signaling with integrin-mediated adhesion and actin cytoskeleton dynamics. It interacts with ITGB1, TLN1, ACTB, and SWIP-1, and is regulated upstream by intracellular Ca2?, SRC, and ITGB1 engagement. Downstream, EFHD1 promotes actin polymerization, focal adhesion turnover, and MAPK signaling. Through these interactions, EFHD1 modulates focal adhesion assembly and disassembly, thereby controlling cell adhesion strength and migration. Representative pathway components such as FAK, SRC, RAC1, and ACTB underscore its central role in transducing adhesion signals.
In HAP1 cells, EFHD1 knockout provides a clean haploid system to study integrin-dependent adhesion without allele compensation. HAP1 cells endogenously express integrins, making the knockout suitable for adhesion, spreading, and migration assays. The haploid nature enhances genetic screening applications, enabling synthetic lethality and modifier screens, and can be employed in forward genetic screens. The leukemic origin also provides a relevant context for studying cancer adhesion biology.
These cells are designed for diverse experimental workflows, including cell adhesion and wound healing assays, immunofluorescence analysis of focal adhesions (e.g., paxillin), western blotting for phospho-FAK, and flow cytometry for integrin surface expression. Live-cell imaging of actin dynamics and high-content screening are also well-suited. Researchers can leverage this model for haploid genetic screens and cancer metastasis studies. For further information, contact Ascent Research.
