The BICD2 Knockout HEK293T Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal cell population derived from the HEK293T line, designed for loss-of-function studies of the BICD2 gene. This polyclonal knockout model introduces disruptive edits across the BICD2 locus, generating a heterogeneous pool of cells with gene disruption, suitable for examining BICD2-dependent cellular processes without clonal selection. The product provides a ready-to-use system for investigating dynein-mediated transport and associated intracellular organization in a human cell context.
HEK293T is a widely used human embryonic kidney cell line exhibiting epithelial morphology, stably expressing the SV40 large T-antigen. This modification allows high-copy episomal replication of plasmids carrying the SV40 origin of replication, enabling efficient transient protein expression and lentivirus production. The parental HEK293 cell line was originally derived from normal embryonic kidney tissue, and the 293T derivative has become a standard workhorse in cell biology laboratories for mechanistic studies and recombinant protein work.
BICD2 encodes a motor adaptor protein that physically links cargoes to the cytoplasmic dynein-dynactin motor complex. In its role as a cargo adaptor, BICD2 is activated by RAB6 GTPase at membranes and regulated by CDK5-mediated phosphorylation. It interacts directly with the dynein heavy chain DYNC1H1 and the dynactin subunit DCTN1/p150Glued, as well as components such as RANBP2, NUP358, and DYNLT1. This interaction network facilitates minus-end-directed transport along microtubules, governing critical processes including Golgi apparatus organization, nuclear envelope positioning, neuronal vesicle trafficking, and mRNA transport granule localization. Disruption of BICD2 therefore impairs recruitment of the motor complex to cargo, leading to aberrant intracellular architecture and defective retrograde trafficking.
In the HEK293T cellular background, knockout of BICD2 presents a valuable model for dissecting the machinery of dynein-dependent vesicular and organelle transport in human epithelial cells. Given the role of HEK293T in producing lentiviral particles, this knockout also offers a platform to study how BICD2-mediated trafficking influences viral protein distribution and virus release. The polyclonal population maintains a representative range of genetic disruptions, allowing researchers to assess overall pathway perturbations while avoiding potential artifacts of clonal outgrowth. Changes in Golgi morphology, nuclear positioning, and localization of associated proteins such as DCTN1 and RAB6A can be readily monitored.
Research applications for these polyclonal knockout cells include mechanistic dissection of dynein motor complex assembly, investigation of Golgi apparatus dynamics and cargo adaptor functions, studies of cell polarity and migration, and analysis of intracellular trafficking pathways relevant to neurodevelopmental disorders and spinal muscular atrophy. Typical assays include immunofluorescence staining for Golgi markers and nuclear position, co-immunoprecipitation of dynein components, live-cell imaging of organelle transport, and downstream gene expression analysis by RT-qPCR. Additionally, the cells support functional assays such as viral protein trafficking and lentivirus production studies. For further details and ordering information, please contact Ascent Research.
