GULP1 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout population in which the GULP1 gene is disrupted, creating a loss-of-function model for investigating GULP1-dependent cellular processes. This product consists of a heterogeneous pool of HAP1 cells carrying diverse CRISPR/Cas9-mediated target-gene disruptions, enabling robust functional studies without single-cell clonal isolation. The pool format minimizes clonal artifacts while preserving the genetic accessibility of the host background, making it suitable for high-throughput screening and pathway analysis where population-level effects of gene disruption are desired.
The host cell line HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia background. These cells exhibit fibroblastoid, adherent morphology and maintain a predominantly haploid karyotype, which greatly facilitates genetic manipulation and phenotypic characterization. The haploid nature simplifies interpretation of knockout phenotypes, as a single targeting event can abolish gene function, allowing efficient generation of loss-of-function models for genes involved in diverse biological pathways.
GULP1 encodes an engulfment adaptor protein that plays a central role in phagocytosis of apoptotic cells and receptor-mediated signaling. Mechanistically, GULP1 binds to the cytoplasmic tail of the low-density lipoprotein receptor-related protein 1 (LRP1), stabilizing the receptor at the cell surface and coupling it to downstream effectors. Through interactions with phosphatidylserine-exposed apoptotic targets and components such as clathrin and ABCA1, GULP1 engages the ELMO/DOCK180/Rac1 GTPase module to drive cytoskeletal rearrangements required for engulfment. Furthermore, GULP1 modulates TGF-beta signaling by influencing SMAD2/3 activation and intersects with the Wnt pathway through beta-catenin regulation. Upstream, LRP1 ligands such as Apolipoprotein E and TGF-beta itself trigger GULP1-dependent cascades, while receptors including BAI1 and TIM4 recognize phosphatidylserine to initiate the engulfment response.
In the HAP1 background, disruption of GULP1 provides a system to dissect how this adaptor coordinates LRP1 trafficking, apoptotic cell clearance, and cross-talk between TGF-beta and Wnt pathways. The polyclonal knockout population enables assessment of bulk cellular responses in assays such as phagocytosis of apoptotic cells, LRP1 surface expression by flow cytometry, and reporter-based quantification of TGF-beta signaling, all while maintaining a near-haploid genetic context that minimizes compensatory allele effects. This model is particularly valuable for probing the tumor suppressor functions of GULP1 and its roles in diseases where defective efferocytosis and receptor dysregulation contribute to pathology.
Researchers can employ these GULP1 knockout cells in a wide range of experimental workflows, including Western blotting and RT-qPCR for verifying loss of GULP1 expression, quantitative engulfment assays to measure clearance of apoptotic corpses, and LRP1 cell surface staining to assess receptor stabilization. The product also supports TGF-beta reporter assays to interrogate downstream SMAD-dependent transcription, as well as studies of cytoskeletal remodeling and Rac1 activation. By eliminating GULP1 function, the model facilitates investigation of molecular mechanisms underlying atherosclerosis, Alzheimer??s disease, and cancer, where GULP1-mediated signaling is dysregulated. For additional details or custom requests, please contact Ascent Research.
