The LIPA Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Raji human B lymphocyte cell line, targeting disruption of the LIPA gene. This polyclonal pool provides a heterogeneous loss-of-function model for studying lysosomal acid lipase (LIPA) in B cells. CRISPR/Cas9-mediated gene disruption ensures efficient abrogation of LIPA expression without clonal bias, enabling robust phenotypic analysis across the mixed population.
The Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma-derived B lymphocyte model widely used in immunology and cancer research. Raji cells, exhibiting mature B cell characteristics and amenability to genetic manipulation, facilitate the study of B cell receptor signaling, apoptosis, and lipid metabolism. Their rapid proliferation and suspension growth support large-scale functional assays and high-throughput screening.
LIPA encodes lysosomal acid lipase, which hydrolyzes cholesteryl esters and triglycerides within lysosomes to release free cholesterol and fatty acids. Its activity is controlled by upstream factors TFEB, PPAR??, LXR, and SREBP2, which regulate lysosomal biogenesis and lipid metabolism. Free cholesterol drives oxysterol production, activating LXR and inducing cholesterol efflux transporters ABCA1 and ABCG1. LIPA interacts with LAMP2, NPC1, NPC2, and SCP-2 in the endolysosomal compartment, forming a cholesterol trafficking network. Disruption of LIPA impairs the LDL receptor?CLDL-cholesterol?Clysosome axis, causing cholesteryl ester and triglyceride accumulation, diminished LXR signaling, defective reverse cholesterol transport, and consequent foam cell formation and inflammation.
In Raji B lymphocytes, LIPA knockout enables investigation of lipid metabolism and immune function interplay. B cells require cholesterol for membrane synthesis and signaling microdomains; impaired LIPA-mediated hydrolysis may disrupt B cell receptor signaling, antigen presentation, and cytokine secretion. The EBV-positive background permits exploration of latent viral persistence under lipotoxic stress, potentially linking lysosomal defects to oncogenic reprogramming. This model is relevant for studying Wolman disease and CESD in hematopoietic cells and for assessing LIPA’s role in B cell malignancies.
Researchers can use this polyclonal knockout model in diverse experimental workflows. Applications include LIPA enzymatic activity assays with fluorogenic substrates, lipid accumulation visualization via filipin or Oil Red O staining, cholesterol efflux measurement, and autophagy monitoring by LC3-II western blotting. The pool suits phenotypic screening of LIPA modulators and enzyme replacement therapy mechanism studies. Lysosomal pH measurements and flow cytometry-based lipid quantification further characterize metabolic disturbances. For additional details, please contact Ascent Research.
