The DNAI2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DNAI2 gene in the HAP1 near-haploid human cell line. This loss-of-function model targets the intermediate chain of the axonemal outer dynein arm, a component critical for generating ciliary and flagellar movement. The polyclonal composition provides a heterogeneous pool of knockout cells, enabling population-level functional studies without the biases of clonal selection.
The HAP1 host cell line is a near-haploid, fibroblast-like adherent cell derived from a male KBM-7 chronic myeloid leukemia patient. It is BCR-ABL positive and retains a mostly haploid karyotype, facilitating straightforward CRISPR/Cas9-mediated gene disruption and functional genomics studies. Under defined culture conditions, HAP1 cells can be differentiated to produce primary cilia, offering a simplified and genetically tractable system for investigating ciliary assembly and motility pathways.
DNAI2 encodes dynein axonemal intermediate chain 2, an integral subunit of the outer dynein arm that interacts with heavy chains DNAH5, DNAI1, and light chains DNALI1, along with docking complex components CCDC151 and CCDC114. Its expression is controlled by ciliogenic transcription factors FOXJ1 and RFX, which act downstream of Notch signaling and Multicilin. CRISPR-mediated knockout disrupts outer dynein arm assembly, resulting in immotile cilia and defective mucociliary clearance. This phenotype mimics primary ciliary dyskinesia type 9 (CILD9) and Kartagener syndrome, affecting ciliary beat frequency and left-right axis determination.
In the HAP1 context, the DNAI2 knockout leverages the cell line??s near-haploid genome to reduce genetic redundancy and streamline genotype?Cphenotype analysis. The polyclonal knockout population encompasses a range of disruptive alleles, making it suitable for biochemical, imaging, and functional assays that survey the collective effects of gene loss. Induction of ciliogenesis in these cells permits examination of ciliary markers such as acetylated tubulin and ARL13B, as well as direct measurement of ciliary beat frequency, providing a robust platform for dissecting outer dynein arm biology.
This product supports diverse research applications including mechanistic studies of primary ciliary dyskinesia, chronic respiratory disease, and infertility related to immotile cilia. It is compatible with assays such as high-speed video microscopy for beat frequency quantification, transmission electron microscopy for axonemal ultrastructure analysis, co-immunoprecipitation to assess dynein complex formation, and RT?qPCR for ciliogenesis markers. The model also facilitates drug screening for mucociliary clearance modulators and functional rescue experiments. For further technical details, please contact Ascent Research.
