The DRAM2 Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HeLa human cervical adenocarcinoma cell line, in which the DRAM2 gene has been disrupted to create a loss-of-function model for studying lysosomal regulation of autophagy and apoptosis. This polyclonal format provides a heterogeneous pool of edited cells, enabling robust functional studies without the clonal bias associated with single-cell isolates. The product is designed for researchers investigating the downstream consequences of DRAM2 ablation in cellular homeostasis and stress responses.
The HeLa cell line is an immortalized human epithelial carcinoma line originating from a cervical adenocarcinoma, famously isolated from Henrietta Lacks in 1951. These cells are HPV18-positive and express the viral oncoproteins E6 and E7, which perturb the p53 and retinoblastoma tumor suppressor pathways, respectively. HeLa cells are a cornerstone of cancer research due to their robust growth, ease of transfection, and extensive characterization, making them an ideal host for generating knockout models to dissect oncogenic signaling and cell death mechanisms.
DRAM2 (DNA damage-regulated autophagy modulator 2) is a lysosomal membrane protein that functions as a critical mediator of autophagy and apoptosis downstream of p53 family members. It is transcriptionally activated by TP53, as well as its homologs TP63 and TP73, particularly in response to genotoxic stress such as ??-irradiation and DNA-damaging agents. Once expressed, DRAM2 localizes to lysosomes and contributes to lysosomal membrane permeabilization, thereby releasing cathepsins and other hydrolases that activate the mitochondrial apoptotic pathway. This process involves interactions with pro-apoptotic BCL2 family members BAX and BAK, and culminates in the activation of caspases CASP3 and CASP9. Concurrently, DRAM2 promotes autophagy by modulating the autophagy machinery, including LC3 lipidation and SQSTM1/p62 turnover, and functionally interacts with ATG5 and ATG7. Thus, DRAM2 serves as a node integrating lysosomal function with cell death and survival pathways.
In the HeLa context, DRAM2 knockout provides a powerful tool to dissect the interplay between lysosomal integrity, autophagy, and apoptosis in a cell line with compromised p53 activity due to HPV18 E6-mediated degradation. This model allows researchers to investigate p53-independent functions of DRAM2 or to restore p53 signaling experimentally to study its full regulatory network. Given the link between autophagy modulation and cancer therapy resistance, this knockout system is particularly valuable for cervical cancer research and for screening compounds that target lysosomal pathways.
The DRAM2 Knockout HeLa Polyclonal Cells are suited for a broad range of experimental applications, including mechanistic studies of autophagy flux using Western blot detection of LC3-II and SQSTM1/p62, apoptosis assays via Annexin V/PI staining and caspase activity measurements, and cell viability assessments under chemotherapeutic or genotoxic stress. Co-immunoprecipitation experiments can map DRAM2’s interaction network, while immunofluorescence microscopy using LAMP1 and DRAM2 antibodies visualizes lysosomal localization. RT-qPCR of DRAM2 and p21 serves as a transcriptional readout of p53 activity, and reporter gene assays link DRAM2 expression to p53-dependent transcription. These applications make the product indispensable for advancing research in autophagy, apoptosis, and cancer biology. For additional technical information, please contact Ascent Research.
