CCDC186 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from near-haploid human HAP1 cells, with targeted disruption of the CCDC186 gene. This loss-of-function model provides a genetically mixed pool of cells, reducing clonal artifacts and enabling robust functional studies. The polyclonal format is ideal for pooled screening, bulk biochemical assays, and quantitative trafficking readouts, serving as a flexible platform to dissect CCDC186 roles in Golgi-related membrane transport.
The HAP1 parental line originates from KBM-7 chronic myelogenous leukemia cells and retains a near-haploid karyotype, making it highly effective for CRISPR-based functional genomics. HAP1 cells are adherent and fibroblast-like, with intact trafficking machinery, allowing detailed studies of membrane dynamics in a simplified genetic background. The haploid state facilitates efficient knockout generation and is widely used for systematic loss-of-function screens.
CCDC186 encodes a coiled-coil protein that functions as a Golgi tethering factor, directly interacting with the TRAPP II complex via TRAPPC10 to mediate vesicle tethering and fusion during intra-Golgi and endosome-to-Golgi retrograde transport. This process is regulated by ARF1 and RAB1 GTPases, which coordinate COPI coat recruitment and SNARE-mediated fusion. Downstream, CCDC186 influences Golgi-enzyme localization and secretory cargo sorting, and associates with C10orf76 and other TRAPP subunits, maintaining organelle homeostasis within a network of COPI and Golgi SNAREs.
In HAP1 cells, CCDC186 knockout disrupts Golgi trafficking, modeling tethering factor dysfunction relevant to neurodevelopmental disorders such as Joubert syndrome. The cells also enable investigation of secretory pathway contributions to cancer and metabolic diseases. This polyclonal knockout population supports analysis of phenotypic heterogeneity and selection of trafficking-impaired subpopulations.
These cells are applicable to immunofluorescence imaging of Golgi markers (GM130, TGN46), live-cell tracking of vesicle transport, and VSV-G ts045 cargo secretion assays. Co-immunoprecipitation validates CCDC186?CTRAPPC10 interactions, while western blotting, electron microscopy, and RNA-seq enable comprehensive pathway analysis. Flow cytometry screens for surface protein expression facilitate drug discovery targeting secretion. For further information and ordering, please contact Ascent Research.
