Description

The PLD3 Knockout HeLa Cell Line is a CRISPR/Cas9-edited knockout cell line in which the PLD3 gene has been disrupted to eliminate PLD3 expression. This stable loss-of-function model enables precise study of PLD3-dependent lysosomal processes without transient gene silencing artifacts. The cell line serves as a robust tool for investigating the molecular consequences of PLD3 deficiency in a broadly used human epithelial host.

The host HeLa cell line is an HPV-18-positive, aneuploid human epithelial line derived from cervical adenocarcinoma. HeLa cells are a staple in cancer research and cell biology due to their robust proliferation and ease of manipulation. Their well-characterized background provides a consistent platform for generating knockout models, and they express essential lysosomal machinery, making them appropriate for examining lysosomal and trafficking pathways disrupted by PLD3 knockout.

PLD3 encodes a lysosomal phospholipase D that hydrolyzes phosphatidylcholine into phosphatidic acid and choline, playing a critical role in phospholipid catabolism and exosome biogenesis. PLD3 activity is regulated transcriptionally by TFEB downstream of mTORC1, and it interacts with sortilin (SORT1), amyloid-beta precursor protein (APP), and CD63. PLD3 deficiency leads to phospholipid accumulation, disrupted endosomal-lysosomal trafficking, and altered exosome cargo sorting, linking it to lysosomal dysfunction and neurodegeneration, particularly late-onset Alzheimer??s disease.

In the HeLa context, PLD3 knockout abolishes its phospholipase activity, leading to aberrant phospholipid accumulation and defective lysosomal function. This results in impaired endolysosomal trafficking and exosome biogenesis, phenocopying aspects of lysosomal storage disorders. The cancer cell background offers a simplified system to investigate cell-autonomous PLD3 functions, independent of neuronal factors, and facilitates high-content screening for modulators of lysosomal lipid metabolism and trafficking.

This knockout cell line supports diverse research applications such as Alzheimer??s disease modeling, lysosomal dysfunction studies, exosome biology, and lipid metabolism research. It is suitable for drug target validation and compound screening using assays like phospholipase D activity measurement, LC-MS lipidomics, LysoTracker staining, immunofluorescence for lysosomal markers, exosome isolation, and immunoblotting. For more information or custom applications, please contact Ascent Research.