The DYNC1LI1 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the human DYNC1LI1 gene has been disrupted to create a loss-of-function model for investigating cytoplasmic dynein-dependent intracellular transport, mitosis, and organelle dynamics. This knockout product is supplied as a heterogeneous pool of gene-edited HEK293T cells, enabling researchers to interrogate the collective effects of DYNC1LI1 deficiency without the constraints of single-cell clonal selection. The polyclonal format preserves population-level variation, making it particularly suitable for phenotypes that benefit from averaged biological effects, such as drug response assays or pooled functional genomic screens.
The host cell line, HEK293T, is an immortalized line derived from human embryonic kidney epithelial cells. These cells were transformed with adenovirus E1A and constitutively express the SV40 large T antigen, conferring high transfectability and robust recombinant protein production capability. As a well-established model, HEK293T cells are routinely utilized for viral packaging, transient and stable gene expression studies, and ligand-receptor interaction analyses. Their rapid growth and adaptability to large-scale culture further enhance their utility in high-throughput screening and biochemical purification workflows.
DYNC1LI1 encodes the light intermediate chain of the cytoplasmic dynein motor complex, a critical component of the minus-end-directed microtubule motor machinery. This protein facilitates the transport of diverse cargoes, including late endosomes, lysosomes, Golgi-derived vesicles, autophagosomes, and the mitotic spindle apparatus. DYNC1LI1 functions within a multimolecular network, interacting directly with the dynein heavy chain (DYNC1H1), intermediate chains (DYNC1I2), and the dynactin complex (e.g., DCTN1). Its activity is modulated by upstream regulators such as LIS1 (PAFAH1B1), NDE1, NDEL1, and the CDK1/cyclin B kinase complex, while adaptor proteins like BICD2 and RAB7 link the motor to specific organelles. Disruption of DYNC1LI1 therefore perturbs multiple trafficking pathways and cell cycle?Cdependent processes.
Within the HEK293T background, the DYNC1LI1 knockout model offers a technically accessible system for mechanistic studies. The cell line??s high transfectability allows facile reintroduction of wild-type or mutant DYNC1LI1 for rescue experiments, enabling precise dissection of structure-function relationships. Loss of this light intermediate chain can be exploited to examine consequences on lysosomal positioning, mitotic spindle orientation, and autophagic flux, all of which are observable through established imaging and biochemical methods. Moreover, the knockout cells provide a relevant platform for validating gene therapy targets and screening small-molecule modulators of the dynein complex.
Typical research applications include Western blotting to confirm DYNC1LI1 depletion, immunofluorescence-based assays for organelle distribution, live-cell imaging of lysosomal movement, and co-immunoprecipitation to assess dynein complex integrity. Additional assays such as flow cytometry for cell cycle profiling, mitotic spindle morphology analysis, and migration/invasion studies can be employed to link DYNC1LI1 function to cell division and motility phenotypes. Transcriptomic profiling by RNA-seq further allows global assessment of pathways affected by the knockout. For additional technical support, custom product requests, or detailed application guidance, please contact Ascent Research.
