The DNASE2 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-mediated polyclonal knockout cell population derived from the NCI-H1299 non-small cell lung cancer line. This loss-of-function model targets DNASE2, a lysosomal endonuclease critical for DNA degradation and innate immune regulation. The polyclonal format provides a heterogeneous genetic disruption suitable for pooled functional analyses without the need for single-cell cloning.
The NCI-H1299 host cell line originates from a human lung adenocarcinoma lymph node metastasis and is p53-null. It is extensively utilized in tumor biology and drug response studies, offering a clinically relevant model for non-small cell lung cancer. Its genetic background facilitates research into metastasis, DNA damage responses, oncogenic signaling, and therapeutic development.
DNASE2 is a lysosomal endonuclease that degrades DNA in acidic conditions, underpinning apoptotic DNA breakdown and immune regulation. Its expression is controlled by the MITF transcription factor and TNF-??. Loss of DNASE2 causes accumulation of self-DNA, which activates the innate immune sensors TLR9 and cGAS-STING, leading to MYD88/NF-kB-dependent production of type I interferons and cytokines such as IFN-?? and IL-6. Lysosomal targeting requires interaction with the mannose-6-phosphate receptor.
In the p53-null NCI-H1299 background, DNASE2 knockout illuminates connections between DNA sensing and tumor immunity. Lung adenocarcinoma cells often display aberrant apoptosis and autophagy, generating DNA substrates for DNASE2. Disruption of DNASE2 may heighten innate immune activation through accumulated DNA, affecting tumor inflammation and responses to therapy. This model thus helps decipher how cancer cells manage immunogenic DNA and evade immune surveillance, especially in the context of defective p53 signaling and genomic instability.
These polyclonal knockout cells are adaptable to numerous research applications, including functional studies of DNASE2-dependent DNA clearance, TLR9/cGAS-STING pathway activation, and autoimmune disease mechanisms such as systemic lupus erythematosus. Typical assays involve Western blotting, RT-qPCR, immunofluorescence, acid DNase activity measurement, ELISA for inflammatory cytokines, and flow cytometry for apoptotic cell uptake. Transcriptomic profiling via RNA-seq and TLR9 reporter assays further elucidate pathway dynamics. The model also supports drug target validation for inflammatory disorders and cancer immunology investigations. For further details or to request a quote, please contact Ascent Research.
