The IFT74 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the IFT74 gene in HAP1 cells. This mixed pool harbors a variety of loss-of-function alleles, providing a robust system for studying IFT74-dependent biology. The polyclonal format ensures high reproducibility for population-level analyses in functional genomics, drug screening, and pathway dissection.
HAP1 is a near-haploid, fibroblast-like cell line derived from KBM-7 chronic myeloid leukemia cells. It carries the BCR-ABL1 translocation, serving as a well-established hematopoietic cancer model. Its near-haploid genome facilitates efficient gene editing, and HAP1 cells can form primary cilia, enabling cilia biology studies in a leukemic context.
IFT74 is a core subunit of the IFT-B complex, essential for anterograde intraflagellar transport and ciliogenesis. It directly interacts with IFT81 and IFT88, and cooperates with kinesin-2 motor KIF3A to deliver structural and signaling components to the cilium tip. Knockout of IFT74 abolishes primary cilium formation, disrupting Hedgehog signaling by impairing SMO and GLI transcription factor activity, as well as affecting Wnt/??-catenin and cell cycle regulation. IFT74 expression is controlled by RFX transcription factors and FOXJ1, and its stability is modulated by AURKA and the ubiquitin-proteasome system. Loss of IFT74 thus dismantles the IFT-B complex, blocks anterograde trafficking, and cripples ciliary signal transduction.
In HAP1 cells, IFT74 knockout results in complete loss of cilia, offering a clean system to investigate cilia-dependent versus -independent mechanisms in BCR-ABL1-driven leukemia. This enables dissection of oncogenic kinase signaling cross-talk with Hedgehog or Wnt pathways by comparing wild-type and IFT74-deficient cells. The near-haploid background further allows efficient introduction of secondary mutations for synthetic lethality screens targeting ciliopathy or leukemia vulnerabilities.
Applications include ciliopathy disease modeling, IFT-B complex assembly analysis, and Hedgehog pathway drug screening. Assays such as immunofluorescence for acetylated ??-tubulin, RT-qPCR of GLI1 and PTCH1, Hedgehog luciferase reporters, Western blotting, and cell migration/invasion assays are readily implemented. The polyclonal format is ideal for pooled genetic interactions or drug synergy studies. For additional information, contact Ascent Research.
