The IFT27 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population designed for targeted disruption of the IFT27 gene in the HAP1 human chronic myeloid leukemia cell line. This loss-of-function model harnesses the endogenous non-homologous end joining repair pathway following Cas9-mediated double-strand breaks to generate a heterogeneous pool of IFT27-null alleles, providing a versatile tool for functional genomics and ciliary biology research without clonal selection.
HAP1 cells, a near-haploid derivative of the KBM-7 chronic myeloid leukemia line, exhibit a predominantly haploid karyotype with chromosome 15 heterogeneity, enabling unambiguous genotype-phenotype correlations in knockout studies. Their growth characteristics and amenability to genetic manipulation make them a widely adopted host for CRISPR/Cas9 screens, signaling assays, and disease modeling, particularly in hematological malignancies and ciliary disorders.
IFT27, a Ras-like small GTPase, heterodimerizes with IFT25 within the IFT complex B to regulate anterograde ciliary trafficking. It transports ciliary building blocks and signaling molecules, including the hedgehog transducer SMO and GTPase ARL13B, and is essential for ciliogenesis. IFT27 promotes nuclear translocation of GLI transcription factors, inducing targets such as GLI1 and PTCH1. It operates within a network of IFT components (IFT172, IFT20) and the BBSome, linking ciliary import to intraflagellar transport.
In the HAP1 model, ablation of IFT27 directly perturbs ciliogenesis and ciliary signal transduction, capturing core defects observed in ciliopathies such as cranioectodermal dysplasia and short-rib polydactyly syndrome. The near-haploid background eliminates confounding effects from residual wild-type alleles, ensuring that observed phenotypes arise from complete loss of IFT27 function. This system enables dissection of hedgehog, GPCR, and other ciliary signaling cascades with high fidelity, and supports the evaluation of small-molecule modulators targeting ciliary trafficking.
Researchers can employ this polyclonal knockout pool in a variety of downstream assays, including immunoblotting for IFT27 protein depletion, immunofluorescence staining of primary cilia with acetylated tubulin and ARL13B, RT-qPCR quantification of GLI1 and PTCH1 transcript levels, automated ciliogenesis scoring, flow cytometry-based phenotypic profiling, and transcriptomic analysis via RNA-seq. These applications position the cell product as a robust platform for investigating ciliary assembly mechanisms, hedgehog pathway dynamics, and the molecular pathology of ciliopathies. For further technical support or custom requests, please contact Ascent Research.
