The DNASE2 Knockout A-549 Polyclonal Cells consist of a polyclonal population of human A-549 lung adenocarcinoma epithelial cells engineered using CRISPR/Cas9 to disrupt the DNASE2 gene. This knockout model eliminates DNASE2 protein expression, enabling functional studies of the lysosomal endonuclease in DNA clearance, innate immune signaling, and programmed cell death.
The A-549 cell line, derived from a lung adenocarcinoma, serves as a model of alveolar type II pneumocytes. It harbors a KRAS G12S activating mutation with wild-type EGFR and p53. This genetic background is widely used to study lung cancer biology and oncogene-driven signaling, making it suitable for investigating how DNASE2 loss impacts KRAS-mutant tumor cells, including potential effects on lysosomal function and inflammatory responses.
DNASE2 encodes a lysosomal endonuclease that hydrolyzes DNA under acidic conditions, preventing accumulation of immunostimulatory DNA. Transcriptionally regulated by TFEB and MITF, the enzyme functions as a homodimer and collaborates with cathepsins. Loss of DNASE2 causes cytosolic leakage of undigested DNA, which triggers the cGAS-STING pathway, leading to IRF3 and NF-kB activation and type I interferon production. Endosomal TLR9 and the AIM2 inflammasome are also engaged, amplifying innate immune signaling. Consequently, DNASE2 knockout establishes a constitutive innate immune activation state driven by endogenous nucleic acids.
In A-549 cells, which express oncogenic KRAS G12S, DNASE2 knockout enables dissection of crosstalk between lysosomal DNA degradation, DNA sensing, and KRAS-driven pathways. This model is relevant to lung adenocarcinoma, where defective clearance of apoptotic DNA may shape the tumor immune microenvironment and influence responses to therapy. It also provides a platform to study mechanisms of type I interferonopathies and autoimmune disorders within an epithelial context.
This polyclonal knockout pool supports applications such as western blotting, RT-qPCR, DNA degradation assays, immunofluorescence, apoptosis analysis, and ELISA for type I interferons. cGAS-STING pathway activation can be assessed by phospho-STING or IRF3 translocation. The model is ideal for studying lysosomal biology, apoptotic DNA clearance, erythropoiesis, autoimmune mechanisms, and drug screening for lysosomal storage diseases or interferonopathies. For additional information, please contact Ascent Research.
