The DPCD Knockout K-562 Polyclonal Cells constitute a heterogeneous population of K-562 cells subjected to CRISPR/Cas9-mediated disruption of the DPCD gene. This pool of polyclonal knockout cells is provided ready-to-use, enabling loss-of-function studies in a familiar hematopoietic cell model. Disruption of the DPCD locus ablates protein expression, thus offering a flexible tool for probing DPCD-dependent cellular mechanisms.
The parental K-562 cell line was established from a 53-year-old female with chronic myelogenous leukemia in blast crisis and is characterized by BCR-ABL positivity and suspension growth. Widely used for CML research, these cells also support large-scale biochemical investigations. Importantly, K-562 cells do not exhibit endogenous ciliogenesis, meaning they lack cilia under standard conditions. This feature provides an exceptional null background for functional studies of ciliary proteins like DPCD, where re-introduction experiments yield unambiguous results.
DPCD encodes a protein critical for motile cilia assembly and function. Its transcription is controlled by master ciliogenic regulators FOXJ1, RFX2, and RFX3. At the post-translational level, DPCD interacts with intraflagellar transport protein IFT88, coiled-coil domain-containing protein CCDC103, and dynein assembly factor DNAAF3. Together with axonemal dynein heavy chain DNAH5 and intermediate chain DNAI1, DPCD contributes to the formation of functional outer and inner dynein arms. These molecular assemblies generate ciliary motility, driving mucociliary clearance and establishing left-right asymmetry during embryogenesis. Consequently, loss-of-function mutations in DPCD manifest as primary ciliary dyskinesia, often accompanied by Kartagener syndrome and situs inversus.
Utilizing K-562 cells for DPCD knockout leverages their non-ciliated phenotype. The polyclonal knockout pool eradicates endogenous ciliary pathways, permitting researchers to ectopically express DPCD variants or to stimulate ciliogenesis through serum depletion or pharmacological intervention. This experimental platform is particularly advantageous for structure?Cfunction analyses, domain mapping, and real-time assessment of ciliary complex assembly. Moreover, the polyclonal nature captures a spectrum of knockout alleles, minimizing clonal bias and increasing the reproducibility of biochemical findings.
The DPCD Knockout K-562 Polyclonal Cells support diverse molecular and cellular assays. They are highly suitable for co-immunoprecipitation experiments to validate interactions with IFT88, CCDC103, or DNAAF3, and for western blotting to confirm DPCD ablation. Upon ciliogenesis induction, the knockout cells enable immunofluorescence staining for ciliary markers (e.g., acetylated tubulin, ARL13B) and RT-qPCR profiling of ciliogenic transcription factors such as FOXJ1 and RFX2. Luciferase reporter assays can further dissect the signaling pathways governing cilia formation. As a research tool, this polyclonal knockout population facilitates high-throughput screens for novel ciliary assembly factors and in-depth biochemical characterization of motile cilia biology. For additional information, please contact Ascent Research.
