The DNASE1L3 Knockout HAP1 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, in which the DNASE1L3 gene has been disrupted. This heterogeneous mixture of DNASE1L3-null cells provides a robust model for studying the functional roles of the secreted endonuclease without the constraints of clonal selection, and is particularly suited for pooled CRISPR screening and bulk functional assays.
The HAP1 host cell line is a near-haploid human cell line (disomic for chromosome 8) derived from the KBM-7 chronic myeloid leukemia (CML) line, originally isolated from a patient in blast crisis. It displays adherent fibroblastoid morphology and carries a p53-null background, rendering it exceptionally useful for CRISPR-based functional genomics and high-throughput screening in a myeloid lineage context.
DNASE1L3 encodes a Ca2?/Mg2?-dependent endonuclease secreted to degrade extracellular DNA from apoptotic cells and neutrophil extracellular traps (NETs). This prevents accumulation of self-DNA that would otherwise activate innate immune sensors like TLR9 and the cGAS-STING pathway. Expression is regulated by TNF-?? and IL-1?? via NF-??B signaling. The enzyme interacts with histones, HMGB1, and DNA-containing immune complexes to facilitate clearance, thereby suppressing type I interferon and anti-DNA autoantibody production. Loss of function leads to persistent cGAS-STING and TLR9 activation, driving autoimmunity.
In the myeloid-derived HAP1 background, knockout of DNASE1L3 offers a genetically tractable platform to interrogate mechanisms of extracellular DNA sensing and innate immune activation. The p53-null status of HAP1 may influence DNA damage response pathways, making this model particularly relevant for studying the interplay between apoptosis, NETosis, and autoimmunity. This knockout population enables investigation of signaling events that drive diseases such as systemic lupus erythematosus, rheumatoid arthritis, and ANCA-associated vasculitis.
This polyclonal knockout cell model is well-suited for a broad range of applications, including DNA degradation assays to measure endonuclease activity, NETosis assays to evaluate NET formation and clearance, and cGAS-STING pathway reporter assays. Standard molecular biology techniques such as Western blotting, RT-qPCR, and anti-dsDNA ELISA can be applied to assess expression changes and autoantibody production. The cells also support drug discovery efforts for lupus and autoimmune disorders, as well as functional genomics screening to identify novel regulators of extracellular DNA clearance. For further details or technical support, please contact Ascent Research.
