The DNHD1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells with targeted disruption of the DNHD1 gene. This heterogeneous knockout pool provides a robust loss-of-function model for studying the axonemal dynein heavy chain DNHD1 without the biases of clonal isolation.
HAP1 is a near-haploid, fibroblast-like cell line derived from KBM-7 chronic myeloid leukemia cells. Its adherent, male phenotype and haploid karyotype simplify knockout generation and interpretation, making it a widely used model for genetic screening, cancer research, and cell signaling studies.
DNHD1 encodes a dynein heavy chain essential for motile cilia and sperm flagella. Its expression is transcriptionally activated by FOXJ1, RFX2, and RFX3. The protein assembles with DNAH5 and DNAI1 into outer dynein arms that generate ciliary beat frequency. Downstream, ciliary motility influences Hedgehog signaling, linking DNHD1 to developmental and homeostatic processes. Thus, knockout disrupts dynein complex formation, ciliary movement, and signal transduction.
In HAP1 cells, DNHD1 knockout recapitulates motile cilia dysfunction found in ciliopathies such as primary ciliary dyskinesia and male infertility. The haploid background permits near-complete loss of function without biallelic targeting. The CML origin additionally allows exploration of cilia?Ccancer signaling crosstalk, while the polyclonal format supports studies of phenotypic variability.
Applications include immunofluorescence for ciliary markers, beat frequency analysis by high-speed microscopy, RT-qPCR for ciliogenesis genes, and western blotting for dynein subunits. Sanger sequencing verifies target disruption. The cells are amenable to compound screening for ciliary modulators or Hedgehog pathway regulators. For further details, contact Ascent Research.
