The DNAH5 Knockout K-562 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population derived from the K-562 chronic myeloid leukemia (CML) cell line. This pool of edited cells harbors targeted gene disruption of the DNAH5 locus, resulting in loss of DNAH5 protein expression. As a polyclonal population, it represents a heterogeneous mixture of cells with various editing outcomes, enabling robust and reproducible functional studies without clonal selection. The product is ideal for researchers investigating DNAH5-dependent mechanisms in a hematopoietic context.
K-562 is a widely used suspension cell line established from the pleural effusion of a 53-year-old female with CML in blast crisis. These cells are characterized by the Philadelphia chromosome, resulting in BCR-ABL1 fusion, and serve as a model for studying leukemia biology and drug responses. Notably, K-562 cells lack ciliated phenotypes, making them a unique host for exploring DNAH5 functions independent of ciliary assembly. The host cell??s genetic background provides a clean system to dissect DNAH5 molecular interactions without confounding ciliary motility effects.
DNAH5 encodes an axonemal dynein heavy chain that constitutes a critical component of the outer dynein arm (ODA) in motile cilia. In ciliated cells, DNAH5 interacts with DNAI1, DNAI2, DNAL1, and TXNDC3 to assemble functional ODA complexes, driving ciliary beat and mucociliary clearance. Transcription of DNAH5 is regulated by FOXJ1 and RFX3, key transcription factors for motile ciliogenesis, and is modulated by NOTCH signaling. Disruption of DNAH5 abrogates ODA motor activity, leading to immotile cilia and impaired airway surface liquid homeostasis, hallmarks of primary ciliary dyskinesia (PCD).
While K-562 cells do not form cilia, the knockout model remains valuable for investigating DNAH5 protein stability, binding partners, and potential non-ciliary functions. The loss of DNAH5 in this leukemic background permits examination of how ODA components behave in the absence of functional axonemes and allows mapping of interaction networks among DNAH5 and its interacting factors without interference from ciliary dynamics. This system can also be used to screen modulators of DNAH5 expression or to study the impact of DNAH5 deficiency on cellular signaling pathways.
This DNAH5 knockout cell pool supports a wide array of research applications, including functional genomics of ciliary motility, validation of CRISPR guide RNA efficiency, and mechanistic studies of PCD. Researchers can employ assays such as western blotting and RT-qPCR to confirm DNAH5 ablation, genomic DNA PCR and T7E1 mismatch detection to assess editing frequency, and immunofluorescence or co-immunoprecipitation to probe protein interactions. For further technical inquiries, please contact Ascent Research.
