The DYNLT3 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from HEK293T human embryonic kidney cells, with targeted disruption of the DYNLT3 gene. This heterogeneous pool enables pooled loss-of-function studies without clonal selection, providing a convenient system for investigating dynein light chain biology in a well-characterized host background. The HEK293T platform supports robust transient and stable expression, making these knockout cells suitable for complementation assays and comparative analyses with wild-type controls to delineate DYNLT3-specific functions.
The parental HEK293T cell line is an immortalized derivative of human embryonic kidney epithelial cells, stably expressing the SV40 large T antigen to enhance episomal replication and protein expression. These cells are widely employed for transient protein production, lentiviral packaging, and genome-scale screening due to their epithelial morphology, rapid growth, and high transfection efficiency. This background is ideal for studying fundamental processes including intracellular trafficking, cytoskeletal dynamics, and organelle biogenesis, and is compatible with a broad range of imaging, biochemical, and functional assays.
DYNLT3 encodes a light chain subunit of the cytoplasmic dynein 1 motor complex, which drives minus-end-directed transport along microtubules. DYNLT3 functions as a cargo-binding adaptor linking dynein to specific cargos and regulating motor activity. It directly interacts with DYNC1H1, intermediate and light intermediate chains, dynactin, and NudE/NudEL. DYNLT3 is modulated by cell cycle machinery, RFX transcription factors driving ciliogenesis, and MAPK signaling. Downstream, DYNLT3 mediates microtubule-based organelle positioning, mitotic spindle orientation, ciliary protein trafficking, and endosomal sorting. Through these interactions, DYNLT3 integrates signaling cues to coordinate dynein-dependent processes critical for cell division, migration, and sensory organelle function.
In the HEK293T context, loss of DYNLT3 disrupts the stability and cargo-recognition capacity of the dynein?Cdynactin complex, leading to defects in retrograde transport, endosomal trafficking, and mitotic spindle assembly. This model enables specific dissection of DYNLT3??s contribution to dynein function, independent of other light chain subunits. The polyclonal nature permits observation of phenotypic variability and dosage effects, which can be further refined through single-cell cloning if desired. Researchers can leverage this system to explore how DYNLT3 loss affects epithelial cell architecture, proliferation, and ciliogenesis, providing insights into dynein-related pathologies including ciliopathies, neurodevelopmental disorders, and cancer.
Applications include live-cell imaging of organelle dynamics, mitotic index and spindle morphology assays, and ciliogenesis quantification. The cells enable co-immunoprecipitation of dynein complexes and cancer cell migration/invasion assays. Western blotting and immunofluorescence can confirm DYNLT3 depletion and monitor dynein subunit localization. These applications support research into the molecular mechanisms underlying ciliopathies, tumor progression, and neurodevelopmental disorders. For additional information and technical support, please contact Ascent Research.
