The DNAL4 Knockout HEK293T Polyclonal Cells comprise a population of human embryonic kidney HEK293T cells engineered with CRISPR/Cas9-mediated disruption of the DNAL4 gene. This polyclonal knockout pool provides a heterogeneous loss-of-function model for investigating the role of the axonemal dynein light chain DNAL4 in ciliary motility. The product eliminates DNAL4 function without requiring clonal isolation, enabling the study of pooled knockout effects across a range of editing outcomes and facilitating robust, statistically powered analyses of ciliary phenotypes.
HEK293T cells are an epithelial cell line derived from human embryonic kidney, stably expressing the SV40 large T antigen. This background confers rapid proliferation and high transfection efficiency, which is advantageous for introducing reporter constructs or performing functional rescue experiments. Although HEK293T cells are non-motile, they can form primary cilia under defined culture conditions and express key components of the dynein arm assembly pathway, including DNAL4, DNAH5, and DNALI1, making them a relevant and tractable model for studying the molecular mechanisms underlying ciliary dynein complex formation.
DNAL4 encodes a light chain of the outer dynein arm, a multi-subunit motor complex essential for ciliary and flagellar motility. The expression of DNAL4 is transcriptionally regulated by the master ciliogenic factors FOXJ1 and RFX2, and its protein product directly interacts with axonemal dynein heavy chains such as DNAH5 and light chain DNALI1 to assemble functional dynein arms. Disruption of DNAL4 impairs the integrity of this complex, leading to defective axonemal dynein arm assembly, reduced ciliary beat frequency, and ciliopathy-related phenotypes, particularly those associated with primary ciliary dyskinesia.
In the HEK293T cellular context, DNAL4 knockout allows the dissection of dynein arm assembly pathways decoupled from motile cilia function, leveraging the cell line’s robust biochemistry and imaging characteristics. The polyclonal knockout format mimics the genetic heterogeneity found in patient populations, enabling dose-dependent or mosaic analyses of DNAL4 loss. This model is particularly suited for monitoring the accumulation and interaction of dynein components via co-immunoprecipitation, as well as for assessing cilia formation and length using markers such as acetylated alpha-tubulin.
This DNAL4 knockout cell product is designed for a broad range of research applications, including primary ciliary dyskinesia disease modeling, mechanistic studies of dynein arm assembly, and functional dissection of cilia-dependent signaling pathways. Typical experimental readouts include western blotting for DNAL4 protein levels, quantitative RT-PCR for DNAL4 transcript disruption, immunofluorescence for cilia markers, and co-immunoprecipitation to examine dynein complex formation. For further details or technical assistance, please contact Ascent Research.
