The ESYT2 Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B lymphocyte line, featuring targeted disruption of the ESYT2 gene. This polyclonal format provides a heterogeneous pool of cells with gene-disrupted alleles, enabling loss-of-function studies without the clonal biases of single-cell isolates. The knockout model is generated by CRISPR/Cas9-mediated genome editing to ablate ESYT2 expression, and it serves as a versatile platform for probing the molecular functions of ESYT2 in a B-cell context.
The parental Raji cell line originates from a Burkitt lymphoma and is Epstein-Barr virus positive, growing in suspension as lymphoblast-like cells. As a B lymphocyte model, Raji cells are widely employed to investigate antibody production, immune surveillance, and signaling in B-cell malignancies. Their transformed phenotype and genetic background make them suitable for studying oncogenic pathways and lipid-dependent processes relevant to lymphoma biology.
ESYT2 is an endoplasmic reticulum (ER)-resident protein that acts as a Ca2+-dependent tethering factor at ER-plasma membrane contact sites. Elevated cytosolic Ca2+ levels, mediated by signals such as epidermal growth factor (EGF) and protein kinase C (PKC), activate ESYT2’s C2 domains, promoting membrane apposition and non-vesicular transfer of glycerolipids. This activity regulates the distribution of phosphatidylinositol 4,5-bisphosphate (PIP2) and actin cytoskeleton organization, thereby influencing lipid homeostasis. ESYT2 functionally interacts with VAPA, VAPB, ESYT1, ESYT3, ORP1, and ORP2, and cooperates with pathway components including OSBPL2, PIKFYVE, and PIP5K1C to maintain phosphoinositide pools and support cellular processes such as proliferation and migration.
In Raji B lymphocytes, disruption of ESYT2 offers a physiologically relevant model to dissect the role of ER-plasma membrane contact sites in immune cell function. Given the involvement of ESYT2 in lipid transfer and Ca2+ signaling, this knockout system enables the examination of how altered membrane dynamics affect antibody production, signal transduction, and malignant transformation. It is particularly valuable for exploring the contribution of non-vesicular lipid transport pathways to the pathogenesis of B-cell lymphomas and for identifying potential therapeutic vulnerabilities.
Researchers can utilize these polyclonal knockout cells in a range of assays, including western blotting and RT-qPCR for confirming target gene disruption, immunofluorescence microscopy to visualize contact site markers, lipid transfer and calcium imaging assays to assess functional consequences, and flow cytometry or cell migration assays to evaluate phenotypic changes. The model facilitates investigations in membrane contact site biology, B-cell lipid metabolism, calcium signaling in lymphocytes, and drug targeting of lipid transfer proteins. For further details or customization, please contact Ascent Research.
