The DPM1 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the HeLa cell line, featuring targeted disruption of the human DPM1 gene. This knockout pool provides a heterogeneous loss-of-function model for studying DPM1-dependent processes without single-cell cloning.
HeLa cells are an immortalized epithelial line from a cervical adenocarcinoma, HPV18-positive, p53 wild-type, and aneuploid. Extensively used in cancer and cell biology research, they offer a robust platform for investigating glycosylation due to their active protein synthesis and well-characterized signaling networks.
DPM1 encodes the catalytic subunit of dolichol-phosphate mannose synthase, which forms a complex with DPM2 and DPM3 on the ER cytoplasmic face. This complex transfers mannose from GDP-mannose to dolichol-phosphate, producing dolichol-phosphate mannose (Dol-P-Man). Dol-P-Man is transported into the ER lumen and serves as the mannosyl donor for N-glycosylation (via ALG1), O-mannosylation (catalyzed by POMT1/2), and GPI anchor synthesis. DPM1 is stabilized by DPM2/DPM3 and influenced by SREBP1/2 and the unfolded protein response. Key downstream targets include alpha-dystroglycan, LAMP1, and GPI-anchored proteins such as CD55.
In HeLa cells, DPM1 knockout disrupts glycosylation, leading to ER stress and altered surface protein expression. This polyclonal model is valuable for studying congenital disorder of glycosylation type Ie (DPM1-CDG) and exploring glycosylation??s role in cancer cell biology. The heterogeneous knockout pool allows assessment of phenotypic variability and modifier effects, while the HeLa background enables mechanistic dissection of glycosylation-dependent processes.
Applications include investigation of CDG type Ie, functional analysis of N- and O-glycosylation and GPI anchor pathways, and screening of glycosylation modulators. Typical assays encompass Western blotting for ICAM-1 and LAMP2, lectin blotting, flow cytometry for CD55 and CD59, UPR gene expression analysis, and mass spectrometry-based glycan profiling. For technical support, contact Ascent Research.
