BET1L Knockout HAP1 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population of the human near-haploid HAP1 cell line, designed to disrupt the BET1L gene. This polyclonal pool provides a loss-of-function model for investigating the role of BET1L in intracellular trafficking. By abolishing BET1L expression, researchers can interrogate its contributions to Golgi homeostasis, vesicular transport, and the secretory pathway. The product is supplied as a heterogeneous population of edited cells, reflecting the polyclonal knockout format, and is suitable for functional studies, genetic screens, and biochemical analyses without clonal selection artifacts.
The HAP1 host cell line originates from the KBM-7 chronic myeloid leukemia lineage and retains a predominantly haploid karyotype, with disomy only for chromosome 8. This near-haploid state minimizes genetic redundancy and facilitates unambiguous genotype?Cphenotype correlations, making HAP1 a workhorse model for functional genomics. The cells are adherent, easily transfectable, and amenable to CRISPR-based editing, enabling robust loss-of-function screening. Their stable genome and rapid growth kinetics support high-throughput assays, and the haploid background simplifies the generation of knockout populations without the need for homozygous editing.
BET1L encodes a Golgi Qb-SNARE protein that forms a cognate trans-SNARE complex with GOSR2, STX5, and YKT6 to mediate COPI vesicle tethering and fusion at the Golgi membrane. This SNAREpin assembly is essential for intra-Golgi retrograde transport and proper maintenance of Golgi architecture. BET1L activity is regulated upstream by ER stress signaling and the XBP1 transcription factor; its loss disrupts COPI-dependent vesicle fusion, leading to Golgi fragmentation, impaired ER-to-Golgi protein transport, and defective protein secretion and glycosylation. Interacting partners such as BET1 and SEC22B further integrate BET1L into the broader ER?CGolgi SNARE network.
In the HAP1 context, BET1L knockout eliminates a critical node in the Golgi fusion machinery, allowing dissection of SNARE-specific trafficking steps against a clean haploid background. This model enables unambiguous assessment of phenotype severity without interference from a second wild-type allele, and it is particularly valuable for teasing apart the contributions of individual Golgi SNAREs. The knockout cells recapitulate hallmark features of Golgi stress, including disrupted Golgi morphology, accumulation of cargo in ER-derived compartments, and activation of the unfolded protein response, providing a tractable system for chemical or genetic rescue experiments.
Typical experimental applications include monitoring ER-to-Golgi transport kinetics using a VSVG-GFP trafficking assay, visualizing Golgi fragmentation by immunofluorescence staining for markers such as GM130 and TGN46, and quantifying secretion defects via Gaussia luciferase reporters. Lectin blotting permits analysis of glycosylation abnormalities, while RT-qPCR for UPR target genes probes the ER stress response. The polyclonal pool is also well-suited for pooled genetic screens aimed at isolating secretory pathway components or modulators of Golgi integrity. For further technical specifications or ordering information, please contact Ascent Research.
