This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from HeLa cells, targeting the DNAH5 gene. The polyclonal pool contains heterogeneous disruptions across the DNAH5 locus, creating a loss-of-function model that avoids single-cell clonal selection. This format provides a robust and reproducible system for studying DNAH5-dependent processes without the constraints of clonal variation, making it suitable for pooled assays and population-level analyses in ciliary biology.
HeLa cells are an aneuploid, immortalized epithelial cell line originally established from a human cervical adenocarcinoma. They are widely employed in cancer biology, cell signaling, and general in vitro research due to their robust growth characteristics, ease of transfection, and extensive legacy as a model system. Although HeLa cells do not constitutively form motile cilia, they can be induced to undergo ciliogenesis through serum starvation or specific differentiation protocols, enabling functional studies of axonemal components like DNAH5.
The DNAH5 gene encodes an axonemal dynein heavy chain that is a structural component of outer dynein arms in motile cilia. DNAH5 protein interacts with other dynein subunits such as DNAI1, DNAH11, and tubulin, and its assembly into axonemal complexes is transcriptionally regulated by FOXJ1 and RFX family transcription factors. In ciliated cells, DNAH5 mediates ATP-dependent microtubule sliding, generating the mechanical force for ciliary beat and driving mucociliary clearance. Disruption of DNAH5 impairs outer dynein arm assembly, thereby compromising ciliary motility and contributing to the molecular pathology of primary ciliary dyskinesia (CILD3) and Kartagener syndrome.
In the HeLa host background, DNAH5 knockout provides a controlled setting to dissect the molecular requirements for outer dynein arm formation and function independently of endogenous ciliogenesis programs. By experimentally triggering cilia formation, researchers can assess whether DNAH5 is required for cilia assembly, maintenance, or motility. The absence of DNAH5 in this tractable cell line facilitates the study of compensatory mechanisms, genetic interactions, and the impact of disease-associated mutations when combined with exogenous expression constructs, offering insights into the broader ciliopathy spectrum.
Typical research applications include western blotting and immunofluorescence for DNAH5 and ciliary markers such as acetylated tubulin and ARL13B to confirm knockout and ciliation status. Ciliogenesis induction coupled with high-speed video microscopy enables motility analysis, while co-immunoprecipitation and mass spectrometry identify DNAH5 binding partners. Transcriptomic profiling via RNA-seq reveals changes in ciliogenesis gene expression upon DNAH5 loss. The polyclonal knockout population is also suitable for small-molecule screens aimed at restoring ciliary function. For additional details and availability, please contact Ascent Research.
