The DNAAF2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the human DNAAF2 gene. This product introduces targeted gene disruption in the widely used HEK293T background, providing a robust tool for population-level analyses of dynein axonemal assembly factor 2 function.
HEK293T cells are adherent, epithelial-like cells derived from human embryonic kidney and express the SV40 large T-antigen, conferring high transfection efficiency and robust protein expression. These features make the line ideal for viral production and biochemical reconstitution studies, enabling the investigation of ciliary protein assembly in a tractable cellular system.
DNAAF2 encodes a cytoplasmic co-chaperone that collaborates with HSPA8 to mediate the folding and preassembly of axonemal dynein complexes. It is critical for the formation of inner and outer dynein arms, which are essential for ciliary motility. Expression of DNAAF2 is controlled by ciliogenic transcription factors FOXJ1, RFX2, RFX3, and MCIDAS, and its protein interacts with assembly partners including DNAAF1, DNAAF3, and DNAJB13. Through these interactions, DNAAF2 facilitates the proper incorporation of dynein heavy and intermediate chains (e.g., DNAH5, DNAH11, DNAI1, DNAI2) into functional motors. Disruption of DNAAF2 thereby impairs cytoplasmic dynein assembly, leading to reduced ciliary beat frequency and compromised mucociliary clearance.
In the HEK293T background, which lacks endogenous motile cilia, the DNAAF2 knockout provides a clean system for dissecting molecular interactions and chaperone functions without interference from native axonemal structures. This model is especially useful for biochemical interaction studies, complementation assays with mutant DNAAF2 variants, and mapping the hierarchy of dynein assembly factors under controlled conditions.
Researchers can employ these cells in immunofluorescence, western blotting, and RT-qPCR workflows to probe dynein component expression and ciliary gene regulation. When coupled with ciliogenesis induction (e.g., FOXJ1 overexpression), the cells become suitable for high-speed video microscopy to measure ciliary beat frequency. Additional applications include drug screening for primary ciliary dyskinesia and evaluating therapies aimed at restoring ciliary function. For further information, please contact Ascent Research.
