The LYPLA1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population derived from the Raji B lymphoblastoid cell line, featuring targeted disruption of the LYPLA1 gene. This product provides a heterogeneous pool of edited cells suitable for functional studies of protein depalmitoylation without the need for single-cell clone isolation.
Raji is an EBV-positive human B lymphocyte line originally established from a Burkitt’s lymphoma patient. These cells retain key B cell features, including surface immunoglobulin expression, robust antigen presentation, and antibody secretion, making them a well-established model for immunological studies and B cell malignancy research. Their high proliferation rate and well-characterized signaling networks support investigation of oncogenic pathways.
LYPLA1 (also known as APT1) encodes a serine hydrolase that catalyzes depalmitoylation??the removal of palmitate from cysteine residues on target proteins. Substrates include the small GTPases HRAS and NRAS, heterotrimeric G protein alpha subunits GNAQ and GNAS, endothelial nitric oxide synthase (eNOS), and Src family kinases. By reversing palmitoylation, LYPLA1 dynamically controls the subcellular localization and signaling activity of these proteins. The enzyme operates within the protein palmitoylation cycle, with regulation by substrate palmitoylation levels, redox conditions, and post-translational modifications. It interacts with palmitoylated substrates and the scaffold protein GIT1. Downstream signaling networks include the RAF1-MAPK1/3 (ERK) cascade via HRAS/NRAS, G protein-coupled receptor pathways through GNAQ/GNAS, and nitric oxide synthesis mediated by eNOS.
In the context of Raji B cells, loss of LYPLA1 disrupts the normal cycling of palmitate on signaling proteins, likely impairing Ras membrane attachment and subsequent MAPK pathway activation. This perturbation is particularly relevant to B cell lymphomagenesis, where oncogenic Ras signaling contributes to uncontrolled proliferation. The knockout also has the potential to alter G protein-dependent signaling and eNOS function, which may influence B cell activation, differentiation, and interaction with the microenvironment. This model thus offers a valuable system to dissect how depalmitoylation controls lymphocyte biology and how its dysregulation may drive Burkitt’s lymphoma pathology.
Researchers can employ these cells to investigate protein palmitoylation in B cell signaling, examine LYPLA1 function in lymphocyte activation, elucidate the role of Ras palmitoylation in B cell lymphoma, and screen chemical inhibitors of depalmitoylation. Representative assay methodologies include the acyl-biotin exchange palmitoylation assay, Ras activity pulldown (Raf-RBD), immunofluorescence microscopy for protein localization, flow cytometry for B cell activation markers, Western blot analysis of phospho-ERK, and XTT-based proliferation measurements. For additional technical support or ordering, please reach out to Ascent Research.
