The GOSR1 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population with targeted disruption of the human GOSR1 gene in HEK293T cells. This loss-of-function model enables investigation of Golgi SNARE protein GS28 function in intra-Golgi retrograde transport and Golgi maintenance, without single-cell cloning. The heterogeneous population retains HEK293T robustness for biochemical, cell biological, and pharmacological studies.
The HEK293T cell line is a widely utilized host system derived from human embryonic kidney cells that stably expresses the SV40 large T antigen. This expression enables high-copy episomal replication of plasmids containing the SV40 origin of replication, making HEK293T cells exceptionally well-suited for recombinant protein overexpression and lentivirus packaging. The cells exhibit adherent epithelial morphology and are maintained in standard culture conditions, providing a reproducible and scalable experimental platform for studying fundamental cellular processes and for producing biological materials.
GOSR1 encodes the Golgi SNARE protein GS28, a Qb SNARE that forms a functional complex with STX5, BET1, and SEC22B to mediate tethering and fusion of COPI vesicles at the cis-Golgi. This SNARE-mediated fusion is essential for retrograde transport of escaped ER proteins and recycling of Golgi enzymes, maintaining Golgi stack organization and glycosylation competency. GOSR1 activity is regulated by NSF, ??-SNAP, Rab1, Sly1 (SCFD1), ARF1, and Golgin-84, and is critical for processing of secretory cargo, glycosyltransferases, and lysosomal enzymes. Disruption of GOSR1 abrogates SNARE assembly, leading to impaired trafficking, Golgi fragmentation, aberrant glycosylation, and potential induction of Golgi stress and apoptosis.
In HEK293T cells, GOSR1 knockout provides a powerful model for dissecting Golgi-dependent trafficking and stress responses. The genetic tractability and rapid growth of HEK293T facilitate generation of polyclonal knockout populations for transient and stable assays. Loss of GOSR1 allows examination of disrupted retrograde transport effects on localization of Golgi markers (GM130, Giantin), secretory cargo trafficking kinetics (VSVG-ts045), and Golgi stress signaling activation. This model is relevant for studying neurodevelopmental disorders, epilepsy, and intellectual disability linked to GOSR1 mutations.
Typical applications include co-immunoprecipitation to assess SNARE complex formation (STX5, BET1, SEC22B); immunofluorescence for Golgi morphology (GM130, Giantin); fluorescent cargo trafficking assays; lectin blotting for glycan analysis; and apoptosis assays for Golgi stress-induced death. These cells are also suitable for high-content screening of pharmacological modulators and electron microscopy for ultrastructural studies. For further information, contact Ascent Research.
