The NOD2 Knockout HL-60 Cell Line is a CRISPR/Cas9-edited knockout cell line originating from the human HL-60 promyelocytic leukemia line. This model provides a loss-of-function system in which NOD2 expression is disrupted, enabling precise studies of intracellular pattern recognition receptor signaling and innate immune processes. The cell line is well-suited for assays investigating NOD2-dependent pathways.
HL-60 cells are a classical model for myeloid differentiation and leukemia, established from a 36-year-old female with acute promyelocytic leukemia. These cells can be induced to differentiate along granulocytic or monocytic/macrophage-like lineages using chemical agents such as DMSO or phorbol esters. This property renders HL-60 useful for investigating myeloid cell functions in both undifferentiated and mature states.
NOD2 functions as a cytosolic receptor for bacterial muramyl dipeptide (MDP). Ligand binding triggers NOD2 self-oligomerization and recruitment of RIPK2 via CARD interactions, which in turn activates the IKK complex and TAK1. Downstream, NF-??B and MAP kinase cascades??including p38, JNK, and ERK??are engaged, leading to transcription of pro-inflammatory cytokines (e.g., TNF, IL-6, IL-8) and antimicrobial peptides such as defensins. NOD2 also interfaces with autophagy through interactions with ATG16L1 and CARD9. Upstream regulators include TNF-?? and TLR agonists, which modulate NOD2 expression, while the receptor itself serves as a critical sensor of microbial wall components.
In the HL-60 background, NOD2 knockout eliminates the primary MDP-sensing pathway, enabling dissection of innate immune signaling in a myeloid progenitor context. Because HL-60 can be driven toward macrophage-like cells??a lineage where NOD2 is highly relevant??this model allows evaluation of receptor-dependent cytokine secretion, NF-??B transcriptional responses, and autophagy induction upon MDP challenge. It is particularly useful for characterizing how NOD2 loss influences differentiation-linked immune functions and for modeling aspects of Crohn??s disease and Blau syndrome.
This knockout cell line supports a wide range of experimental approaches, including western blotting, RT-qPCR, ELISA, NF-??B reporter assays, and MDP stimulation studies to quantify pathway activation. It is also applicable in co-immunoprecipitation of NOD2?CRIPK2 complexes, phospho-signaling analysis of MAPK members, and autophagy flux monitoring. The model facilitates pharmacological testing of RIPK2 inhibitors and functional rescue with disease-associated NOD2 variants. For further inquiries, please contact Ascent Research.
