The AZI2 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HAP1 cell line, engineered to disrupt the AZI2 gene. This product provides a heterogeneous loss-of-function model that eliminates AZI2 protein expression across a mixed pool of edited cells, enabling robust analysis of AZI2-dependent signaling without clonal artifacts.
The host HAP1 cell line is a near-haploid human cell line originally derived from a male chronic myeloid leukemia (CML) patient and exhibits adherent, fibroblast-like morphology. Its haploid genome simplifies gene editing and facilitates the study of loss-of-function phenotypes in a clean genetic background. While originating from a hematopoietic malignancy, HAP1 cells retain functional innate immune signaling pathways, making them a valuable platform for dissecting antiviral and inflammatory responses.
AZI2 (also known as NAP1) functions as a critical adaptor protein that scaffolds pattern recognition receptors to downstream kinases. Following stimulation by viral RNA, poly(I:C), or lipopolysaccharide, AZI2 bridges activated TLR3 and RIG-I/MDA5 to the kinases TBK1 and IKBKE, interacting with adaptors TRIF (TICAM1) and MAVS, as well as TRAF3 and TANK. This assembly promotes phosphorylation of transcription factors IRF3 and IRF7, and activates NF-??B, leading to the transcriptional induction of type I interferons (IFNB1, IFNA) and pro-inflammatory cytokines (IL-6, TNF).
Disruption of AZI2 in the HAP1 haploid background creates a powerful model for deciphering its role in innate immune signaling. The absence of functional AZI2 impairs signal transduction from TLR3 and RIG-I-like receptors, resulting in attenuated activation of TBK1, IRF3, and NF-??B. This knockout model allows researchers to map pathway components and assess the impact of AZI2 loss on antiviral and inflammatory responses without interference from a second allele, making it particularly suitable for high-throughput genetic screens and detailed mechanistic studies.
This polyclonal knockout population is ideal for a range of experimental applications. Users can employ viral infection models with Sendai virus or HSV-1 to study AZI2-dependent antiviral responses, luciferase reporter assays to monitor NF-??B and ISRE activity, and immunoblotting to detect phosphorylated TBK1 and IRF3. Additional techniques include RT-qPCR for IFNB1 and cytokine expression, ELISA for secreted IL-6 and TNF, co-immunoprecipitation to analyze AZI2-TBK1 complexes, RNA-seq for transcriptome-wide analysis, and immunofluorescence to visualize IRF3 nuclear translocation. The model supports investigations into innate immune regulation, autoimmunity, and the intersection of inflammation and cancer. For further information on this product or custom gene-editing services, please contact Ascent Research.
