The DNASE1L1 Knockout HEK293T Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T human embryonic kidney cells, designed to disrupt the DNASE1L1 gene. This gene encodes a secretory deoxyribonuclease that cleaves DNA primarily in apoptotic cells, promoting chromatin breakdown and clearance of cellular debris. The polyclonal population comprises a heterogeneous mixture of edited alleles, providing a robust loss-of-function model without the need for clonal selection, and is intended for investigations into DNASE1L1-mediated DNA cleavage and its roles in apoptosis and immune regulation.
HEK293T cells, the host line for this knockout model, originate from HEK293 cells transformed with sheared adenovirus 5 DNA and stably express the SV40 large T antigen, facilitating episomal replication of SV40 origin-containing plasmids. This feature, along with high transfectability and rapid growth, makes HEK293T cells a workhorse for recombinant protein expression and viral production. Their well-characterized proteome and compatibility with diverse CRISPR delivery methods provide an ideal background for disrupting genes involved in nucleic acid metabolism and cell death pathways.
DNASE1L1 acts downstream of apoptotic signals to exert Ca2+/Mg2+-dependent endonuclease activity on chromatin, complementing intracellular DNases such as CAD/DFF40 and DNASE2 to ensure complete DNA degradation. Representative pathway components include DNASE1, DNASE2, CAD/DFF40, and caspases, with DNASE1L1 mediating DNA cleavage and apoptotic body degradation. Its secretion during apoptosis prevents accumulation of immunogenic self-DNA, thereby modulating innate immune responses, while precise upstream regulatory mechanisms remain incompletely defined.
In the HEK293T context, disruption of DNASE1L1 offers a tractable human cell model for dissecting its contribution to apoptotic DNA fragmentation and extracellular DNA clearance. The high transfection efficiency enables complementation studies with wild-type or mutant DNASE1L1, and the polyclonal population captures a range of disruption efficiencies, useful for dose-response assays such as DNA fragmentation and TUNEL. This model also facilitates exploration of how defective DNA clearance may activate innate immune pathways.
These polyclonal knockout cells are applied in apoptosis research using DNA fragmentation and TUNEL assays, and in autoimmune disease modeling, particularly systemic lupus erythematosus, to study links between impaired DNA degradation and anti-DNA autoantibodies via Western blotting and RT-qPCR. They serve as a DNASE1L1-null background for DNase activity assays and reconstitution experiments, and support screening for modulators of DNASE1L1 function. By enabling dissection of nucleolytic pathways, the cells also aid in investigating crosstalk between apoptotic nucleases and DNA-sensing innate immune sensors. For further information, please contact Ascent Research.
