The EFCAB7 Knockout HEK293T Polyclonal Cells product comprises a polyclonal population of HEK293T cells with CRISPR/Cas9-mediated disruption of the EFCAB7 gene. This loss-of-function model is designed for investigating EFCAB7’s roles in ciliary assembly and calcium-dependent signaling. The heterogeneous knockout pool facilitates robust functional studies without clonal selection bias, making it suitable for initial screening, pathway analysis, and rescue experiments.
HEK293T cells are a derivative of the HEK293 embryonic kidney epithelial line, transformed with adenovirus type 5 DNA and expressing SV40 large T antigen for enhanced protein expression. They retain epithelial morphology and the ability to form primary cilia, making them a tractable model for ciliogenesis studies. Their high transfection efficiency, rapid growth, and well-characterized signaling networks have established them as a standard host for genetic perturbation experiments.
EFCAB7 encodes a calcium-binding protein characterized by EF-hand domains, and is critical for the assembly and function of motile cilia and flagella. Mechanistically, EFCAB7 operates downstream of key ciliogenic transcription factors RFX and FOXJ1, which regulate its expression. The protein interacts directly with calmodulin and various members of the CFAP (cilia- and flagella-associated protein) family, and is integrated within the intraflagellar transport (IFT) machinery, associating with IFT-A, IFT-B, and the BBSome complex. Through these interactions, EFCAB7 promotes the proper organization of ciliary axonemal components and flagellar motility proteins, likely via calcium-mediated structural remodeling. Consequently, disruption of EFCAB7 results in defective cilium formation and flagellar architecture, recapitulating aspects of ciliopathy phenotypes.
In the HEK293T background, EFCAB7 knockout provides a physiologically relevant epithelial system to dissect ciliary assembly and calcium signaling within organelles. Despite their kidney origin, HEK293T cells form primary cilia under serum starvation, enabling direct visualization and quantification of ciliogenesis defects. Loss of EFCAB7 impairs axonemal structure and disrupts intraflagellar transport, making it a valuable model for spermatogenic failure and other cilia-related disorders. The polyclonal nature further allows examination of heterogeneous functional outcomes, mirroring the variable expressivity of ciliopathies.
These EFCAB7 knockout polyclonal cells are ideally suited for immunofluorescence detection of ciliary markers such as acetylated tubulin and ARL13B, Western blotting to confirm loss of EFCAB7 protein, and RT-qPCR profiling of downstream ciliary gene expression. Calcium imaging assays can be employed to investigate disruption of intracellular calcium dynamics. Applications include genetic rescue experiments and small-molecule screening to identify modulators of ciliogenesis and calcium signaling. For further information, please contact Ascent Research.
