DPCD Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the DPCD gene has been disrupted in the human HAP1 cell line. This loss-of-function model is designed to facilitate investigations into the role of DPCD in ciliary motility and primary ciliary dyskinesia (PCD). The polyclonal composition provides a heterogeneous pool of knockout cells, enabling population-level analyses of DPCD deficiency without the bias of clonal selection.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. It is adherent and male, with a near-haploid karyotype that facilitates CRISPR/Cas9-mediated gene disruption and functional genomic screening. Because of its simplified genetic background, HAP1 serves as an effective model for fundamental studies of ciliary protein complexes and N-DRC biology.
DPCD encodes a core component of the nexin-dynein regulatory complex (N-DRC) located in the ciliary axoneme, where it is essential for proper ciliary motility. The DPCD protein interacts directly with other N-DRC subunits including DRC1, DRC3, DRC4, DRC5, and DRC7, and connects to the outer and inner dynein arms and radial spoke proteins. Transcriptional regulation of DPCD is mediated by the RFX family (RFX1, RFX2, RFX3) and FOXJ1, master regulators of ciliogenesis. Loss of DPCD disrupts the N-DRC??s ability to modulate dynein activity, leading to uncoordinated ciliary beating and the clinical manifestations of PCD, such as situs inversus and recurrent respiratory infections. Important downstream effectors affected by DPCD disruption include DNAH5 and DNAI1, key dynein proteins.
Within the near-haploid HAP1 background, polyclonal DPCD knockout generates a powerful loss-of-function tool for investigating N-DRC biology. The haploid genome facilitates complete gene disruption, enabling clear dissection of DPCD??s role in dynein regulatory complex assembly and motor protein function. This model is particularly useful for biochemical and imaging studies aimed at understanding how DPCD deficiency alters N-DRC architecture and protein interactions, providing insights into the molecular pathology of PCD.
The DPCD Knockout HAP1 Polyclonal Cells support a wide array of experimental techniques, including immunofluorescence microscopy to assess localization of N-DRC components, high-resolution video microscopy for ciliary beat frequency analysis, and co-immunoprecipitation to map N-DRC protein interactions. Additional applications include western blotting for protein expression profiling, RT-qPCR to measure ciliogenesis-related gene expression, and air-liquid interface cultures to model mucociliary clearance defects. These applications make the cell population a valuable asset for both basic ciliary biology research and translational studies targeting primary ciliary dyskinesia. For further information, please contact Ascent Research.
