The GRAMD2A Knockout HeLa Polyclonal Cells represent a heterogeneous pool of CRISPR/Cas9-edited HeLa cells carrying targeted gene disruption of GRAMD2A. This polyclonal knockout model facilitates reproducible loss-of-function studies of GRAMD2A-dependent cholesterol trafficking without clonal selection artifacts.
HeLa is an HPV18-positive human cervical epithelial adenocarcinoma cell line, originally derived from Henrietta Lacks, and is a foundational model in cancer biology, virology, and cell signaling research. Its well-characterized lipid metabolism pathways make it an ideal host for investigating intracellular cholesterol trafficking and the regulation of cholesterol homeostasis.
GRAMD2A encodes a sterol-sensing protein that shuttles cholesterol from the plasma membrane to the ER at membrane contact sites. Its GRAM domain detects sterol levels and facilitates interaction with VAP proteins, thereby controlling the proteolytic cleavage of SREBP2. This transcription factor then regulates expression of cholesterol-related genes such as LDLR and HMGCR. Upstream inputs include insulin/IGF-1 signaling and LXR agonists, while GRAMD2A directly binds intracellular cholesterol, membrane phospholipids, and ACAT. Knockout results in impaired ER cholesterol delivery, diminished SREBP processing, and dysregulation of LDL receptor expression, cholesterol esterification, and ER cholesterol accumulation.
In HeLa cancer cells, rapid proliferation demands high cholesterol levels for membrane biogenesis. GRAMD2A disruption perturbs spatial cholesterol distribution, likely affecting membrane fluidity, lipid raft integrity, and associated signaling cascades. This model enables dissection of how aberrant cholesterol trafficking impacts oncogenic signaling, apoptosis, and metabolic adaptations in cervical adenocarcinoma. This model thus provides a valuable system to explore the intersection of lipid metabolism and cancer, as well as potential metabolic disease mechanisms.
These polyclonal knockout cells are suited for fluorescent cholesterol trafficking assays, SREBP luciferase reporter measurement, immunoblotting of SREBP2 cleavage, qRT-PCR of target genes, cellular cholesterol quantification, and immunofluorescence of ER/plasma membrane markers. Lipidomics profiling can further reveal global lipid changes. This model supports drug target validation and ER-plasma membrane contact site investigations. For further technical information, please contact Ascent Research.
