The DNAAF5 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell pool targeting the DNAAF5 gene in the human near-haploid HAP1 cell line. DNAAF5 encodes a cytoplasmic protein essential for the preassembly of axonemal dynein arms, multi-subunit motor complexes that drive ciliary motility. Disruption of DNAAF5 impairs dynein arm formation, leading to immotile cilia and primary ciliary dyskinesia (PCD). This knockout model enables detailed investigation of ciliogenesis and ciliopathy mechanisms.
The HAP1 cell line is a near-haploid derivative of the male-derived KBM-7 chronic myeloid leukemia line, predominantly haploid except for a disomic chromosome 15. It is BCR-ABL1 positive, retaining leukemic signaling pathways, and its haploidy eliminates the complicating effects of a second allele, making it highly suitable for functional genomics, genetic screening, and targeted gene disruption studies.
DNAAF5 functions as a cytoplasmic co-chaperone that cooperates with DNAAF2 (KTU), DNAAF3, HSP70, and HSP90 to fold and assemble axonemal dynein heavy chains (DNAH5, DNAH11) and intermediate chains (DNAI1, DNAI2). Its expression is activated by the transcription factors FOXJ1, RFX2, and RFX3, central to motile ciliogenesis. After preassembly, dynein arms are transported to ciliary axonemes to generate microtubule sliding force for ciliary beat. DNAAF5 deficiency blocks this process, resulting in loss of motility.
In the HAP1 background, DNAAF5 knockout creates a defined loss-of-function system without allelic interference, ideal for dissecting dynein assembly pathways. The polyclonal population averages multiple CRISPR edits, enabling bulk biochemical and functional readouts. This model is particularly valuable for exploring connections between hematological signaling and ciliary biology, as well as for unbiased screens seeking modifiers of dynein arm assembly or ciliary restorers.
Researchers can perform ciliary beat frequency analysis, immunofluorescence localization of dynein arm components, western blotting for DNAAF5 and its chaperone partners, and RT-qPCR quantification of ciliogenesis markers. The haploid platform supports high-throughput phenotypic screens and drug discovery programs against ciliopathies such as primary ciliary dyskinesia and Kartagener syndrome. For technical inquiries and ordering, please consult Ascent Research.
