The DYNC2H1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the DYNC2H1 gene. This product provides a heterogeneous pool of HAP1 cells carrying disruptive edits in DYNC2H1, enabling researchers to investigate gene function without the need for single-cell cloning. The polyclonal format preserves genetic diversity and reduces the risk of off-target clonal effects, making it suitable for pooled screening and robust phenotypic analyses.
The host cell line HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myelogenous leukemia line. These cells exhibit a fibroblast-like adherent morphology and a male karyotype, and their haploid state simplifies genetic manipulation by ensuring that a single allelic disruption can yield a functional knockout. HAP1 cells are widely employed in haploid genetic screens, knockout model generation, and functional genomics due to their efficient gene editing and well-characterized growth characteristics.
DYNC2H1 encodes the heavy chain of cytoplasmic dynein-2, the motor for retrograde intraflagellar transport (IFT) in primary cilia. The dynein-2 complex, comprising DYNC2H1 alongside DYNC2LI1, WDR34, and WDR60, moves IFT particles and ciliary cargo from the tip to the base. Its activity is regulated by Aurora A kinase and CDK-like kinases, and it works in concert with IFT-A and IFT-B adaptor complexes. Knockout of DYNC2H1 halts retrograde transport, leading to defective ciliary assembly and disrupted Hedgehog signaling: GLI transcription factors fail to be properly processed, Smoothened and ciliary GPCRs are mislocalized, and downstream target gene expression is altered.
In the HAP1 background, knockout of DYNC2H1 ablates dynein-2-mediated retrograde transport, resulting in stunted or absent primary cilia and aberrant Hedgehog signaling. Because HAP1 cells are haploid, the polyclonal knockout population provides a near-complete loss of function, enabling clear phenotype identification in ciliogenesis assays and signaling readouts. This model recapitulates molecular defects observed in ciliopathies such as short-rib thoracic dysplasia and Jeune syndrome, and it offers a scalable platform for mechanistic dissection of IFT-related disorders without the complexity of diploid systems.
Researchers can employ this knockout pool in ciliopathy disease modeling, Hedgehog pathway analysis, and drug screening. Representative assays include immunofluorescence for ciliary markers (ARL13B, acetylated tubulin), Western blotting for IFT proteins, live-cell imaging of IFT, qPCR for Hedgehog targets (GLI1, PTCH1), and ciliogenesis assays. The polyclonal format supports pooled functional genomics and high-content screening. For ordering or technical support, contact Ascent Research.
