The NLN Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, engineered to disrupt the NLN gene encoding the zinc metallopeptidase neurolysin. This gene-edited model provides a robust loss-of-function system for studying neurolysin-dependent peptide hormone processing and neurotensin signaling in a B-cell context, with direct relevance to neuro-immune interactions and B-cell malignancy research. The polyclonal product format ensures representation of a broad spectrum of gene edits, making it suitable for diverse functional assays.
The host Raji cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma B lymphocyte line originally established from a male patient. These cells express classic B-cell surface markers and are widely used as a model for B-cell malignancies, offering a well-characterized platform for investigating signal transduction, proliferation, and immune modulation. The NLN knockout in this background allows precise dissection of neuropeptide-mediated signaling effects within a malignant B-cell environment.
Neurolysin functions as a zinc-dependent endopeptidase that cleaves neurotensin and other vasoactive peptides, including bradykinin, substance P, and angiotensin I. CRISPR/Cas9-mediated disruption of NLN eliminates this proteolytic activity, resulting in accumulation of intact neurotensin and sustained engagement of neurotensin receptors NTSR1, NTSR2, and sortilin/NTSR3. Ligand binding triggers G??q-dependent activation of phospholipase C (PLC), leading to inositol trisphosphate-mediated calcium release and downstream phosphorylation of ERK1/2. This signaling cascade is modulated by upstream factors such as inflammatory cytokines and cellular stress, positioning neurolysin as a critical regulator at the intersection of peptide hormone metabolism and MAPK pathway activation.
In Raji B lymphocytes, the loss of neurolysin activity decouples neurotensin inactivation, potentially enhancing calcium and ERK-dependent cellular responses including proliferation, cytokine secretion, and survival. This cellular model is uniquely suited to investigate how neuropeptide signals influence B-cell function in both physiological immunity and pathological states such as lymphoma progression. Moreover, it provides a tool to study neuro-immune crosstalk mechanisms underlying pain hypersensitivity and metabolic disorders where neurotensin signaling is implicated.
Typical research applications include Western blotting and RT-qPCR to confirm NLN gene disruption, neurotensin degradation assays to quantify enzymatic activity loss, intracellular calcium flux measurements, and phospho-ERK analysis to monitor downstream signaling. Functional assays such as B-cell proliferation, cytokine ELISA, and flow cytometry for surface marker retention further enable comprehensive phenotypic characterization. This product is also amenable to high-throughput screening for neurolysin inhibitors or modulators of neurotensin receptor pathways in B-cell malignancies. For further information or to discuss custom projects, please contact Ascent Research.
