The EFCAB7 Knockout HAP1 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout population of the near-haploid human HAP1 cell line with targeted disruption of the EFCAB7 gene. This polyclonal knockout cell pool provides a loss-of-function model to study EFCAB7, an EF-hand calcium-binding protein that plays a pivotal role in ciliary trafficking and hedgehog signal transduction. The polyclonal format yields a heterogeneous mixture of edited alleles, making it suitable for functional genomics screening and phenotype validation without the clonal biases inherent to single-cell-derived knockout lines.
The HAP1 parental cell line is a near-haploid human chronic myeloid leukemia model originating from the KBM-7 cell line. HAP1 cells are male, adherent, and carry a mutation in the TP53 tumor suppressor gene. Their near-haploid state, retaining approximately 25% haploid chromosomes, facilitates unambiguous genotype-phenotype correlations because a single CRISPR-induced mutation can generate a complete loss-of-function effect. This genetic simplicity, combined with the cancer-relevant TP53 mutation, makes HAP1 a favoured host for knockout screens and mechanistic studies in signal transduction research.
EFCAB7 encodes an EF-hand calcium-binding protein that acts as a scaffolding adaptor in the primary cilium. It forms a complex with IQCE and interacts with intraflagellar transport complex A (IFT-A) components IFT122 and IFT140. This EFCAB7-IQCE-IFT-A module mediates the ciliary trafficking of Smoothened (SMO), the key transducer of the hedgehog pathway. Upon SHH binding to Patched-1 (PTCH1), SMO is de-repressed and traffics to the cilium; EFCAB7 facilitates this transport, enabling activation of GLI transcription factors and expression of hedgehog target genes. Therefore, EFCAB7 functions downstream of SHH and calcium signals and upstream of GLI-mediated transcription, linking extracellular cues to the transcriptional output of the pathway.
In HAP1 cells, the haploid genetic background makes EFCAB7 knockout a potent model for studying ciliary trafficking and hedgehog signaling. Loss of EFCAB7 is expected to directly impair SMO localization to cilia and suppress GLI activity, with no confounding effects from a wild-type allele. The TP53 mutation in HAP1 further deregulates cell cycle checkpoints, offering a cancer-relevant context to explore hedgehog-driven proliferation. This combination enables precise dissection of the EFCAB7-IQCE-IFT-A axis and its role in ciliogenesis and signal transduction.
The EFCAB7 knockout HAP1 polyclonal population is ideally suited for a range of experimental approaches. Immunofluorescence staining can be used to assess SMO and IFT component localization within the primary cilium, while Western blotting and RT-qPCR quantify changes in hedgehog pathway protein and mRNA levels, including GLI family transcription factors. GLI-responsive luciferase reporter assays provide a functional readout of pathway activity, and cilia formation assays can determine the impact on ciliogenesis. The polyclonal nature of the cells also enables pooled CRISPR screens, dose-response profiling with hedgehog pathway modulators, and studies of synthetic lethality in ciliopathy-relevant conditions. For further information or to request a quote, please contact Ascent Research.
