LRP2 Knockout IPEC-J2 Cell Line

The LRP2 Knockout IPEC-J2 Cell Line is a CRISPR/Cas9-edited knockout cell line with targeted disruption of the LRP2 gene in the IPEC-J2 porcine jejunal epithelial cell background. This loss-of-function model eliminates functional expression of megalin, the large endocytic receptor encoded by LRP2, providing a defined system for studying receptor-mediated endocytosis and ligand uptake in intestinal epithelial cells.

IPEC-J2 cells are a non-transformed, immortalized cell line derived from neonatal piglet jejunal epithelium. They retain key characteristics of primary intestinal epithelial cells, including formation of polarized monolayers with tight junctions, expression of brush border enzymes, and the capacity for vectorial transport. IPEC-J2 cells are widely used as a physiologically relevant model for investigating intestinal barrier function, nutrient absorption, and innate immune responses.

The LRP2 gene encodes megalin, a transmembrane endocytic receptor of the LDL receptor family that is abundantly expressed on the apical surface of absorptive epithelia. Megalin interacts with cubilin and the chaperone RAP (LRPAP1) to mediate internalization of multiple ligands, including the intrinsic factor-vitamin B12 complex, retinol-binding protein, and vitamin D-binding protein. Transcriptional regulation of LRP2 is governed by upstream factors including VDR, retinoic acid, PPAR??, and HNF4??. Following ligand uptake, megalin facilitates delivery of vitamins to lysosomes, supports retinol utilization, and modulates intracellular signaling through MAPK cascades and synthesis of 1,25(OH)2D3. Additionally, megalin participates in morphogen trafficking, binding SHH and BMP4 to influence epithelial signaling networks.

Disruption of LRP2 in IPEC-J2 cells provides a powerful tool for dissecting the role of megalin-mediated endocytosis in intestinal nutrient handling. The knockout line enables investigation of vitamin B12 and retinol absorption, lipoprotein metabolism, and receptor cycling under controlled in vitro conditions. Because megalin dysfunction is linked to Donnai-Barrow syndrome??a disorder featuring renal tubular resorption defects and neural tube defects??this knockout cell model also offers insight into the intestinal component of systemic receptoropathies. Moreover, the IPEC-J2 LRP2 knockout background allows assessment of megalin as a route for targeted drug delivery across the intestinal epithelium.

This cell line is suitable for a broad range of experimental applications, including western blotting and RT-qPCR to confirm LRP2 disruption, immunofluorescence to examine megalin localization, and endocytosis assays using fluorescently labeled ligands such as vitamin B12, LDL, or retinol-binding protein. Additional uses include co-immunoprecipitation to probe interactions with cubilin or RAP, and transepithelial transport studies to evaluate macromolecular trafficking. It supports research into intestinal absorption mechanisms, receptor-mediated signaling, and disease models of megalin deficiency. For more information, please contact Ascent Research.