The DYNLT1 Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal cell population carrying targeted disruption of the DYNLT1 gene. This product provides a loss-of-function model in HAP1 cells for investigating the cytoplasmic dynein light chain Tctex-type 1 subunit. The polyclonal knockout population preserves heterogeneous editing outcomes across the cell pool, enabling robust functional studies without single-cell clonal isolation. DYNLT1 disruption is achieved through CRISPR/Cas9-mediated gene targeting, generating a versatile tool for examining dynein-dependent intracellular processes.
The HAP1 host cell line is a near-haploid human myeloid leukemia cell line derived from KBM-7 chronic myeloid leukemia cells. HAP1 cells are BCR-ABL positive and retain a stable near-haploid karyotype, making them exceptionally suited for functional genomics applications, including haploid genetic screens and gene-trap mutagenesis. Their haploid state simplifies the generation of complete gene knockouts and facilitates unambiguous genotype?Cphenotype correlations, enabling efficient dissection of gene function in a human cellular context.
DYNLT1 encodes a 14-kDa light chain subunit of the cytoplasmic dynein motor complex, which is essential for retrograde transport of organelles, vesicles, and signaling endosomes along microtubules. This subunit mediates cargo binding through direct interactions with dynein intermediate chain (DYNC1I) and the LC8 family of light chains, and it associates with cargo adaptors such as BICD2 and Hook3. DYNLT1 also interacts with the BBSome, rhodopsin, and DISC1, linking dynein to ciliary transport and neuronal functions. Its activity is regulated by FOXJ1 during ciliogenesis and by CDK1-mediated phosphorylation during mitosis. Downstream, DYNLT1-dependent dynein function is critical for signaling endosome trafficking, proper mitotic spindle positioning, and centrosomal protein distribution, thereby integrating mechanical and signaling pathways.
In the HAP1 near-haploid background, DYNLT1 knockout disrupts cytoplasmic dynein motor activity, providing a clean cellular model to examine consequences of impaired retrograde transport. This system is particularly relevant for studying mitotic defects, primary cilium dysfunction, and aberrant vesicle trafficking without interference from a second wild-type allele. The knockout phenotype can be assessed under conditions that challenge microtubule-dependent processes, offering insights into dynein-related pathologies such as neurodegenerative diseases and ciliopathies.
This DYNLT1 knockout cell population is designed for diverse research applications, including functional genomics, drug target validation, and mechanistic studies of intracellular transport. Typical assays include immunofluorescence to assess dynein localization, live-cell imaging of organelle motility, cell cycle analysis by flow cytometry, co-immunoprecipitation to probe dynein complex integrity, and migration assays to evaluate dynein-dependent cell motility. It also enables genetic interaction screens and evaluation of candidate inhibitors. For further details, please contact Ascent Research.
