DRAM1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line, engineered to disrupt the expression of the DRAM1 gene. This product serves as a loss-of-function model for studying autophagy, apoptosis, and p53 signaling pathways. Unlike clonal lines, the polyclonal format represents a pool of edited cells, avoiding artifacts from single-cell selection while providing robust gene disruption for functional studies.
The HAP1 host cell line, originating from the KBM-7 chronic myeloid leukemia line, is near-haploid, which greatly facilitates the generation of complete gene knockouts. Its genetic stability and ease of manipulation make it an ideal platform for genetic screens and functional genomics research. HAP1 cells retain key signaling pathways relevant to cancer biology, including intact p53 and autophagy machinery, enabling physiologically meaningful readouts for DRAM1 functional analysis.
DRAM1 encodes a lysosomal membrane protein that is transcriptionally activated by TP53 in response to DNA damage, nutrient deprivation, and mTOR signaling inhibition. DRAM1 functions downstream of p53 to stimulate autophagy by promoting LC3 lipidation, p62/SQSTM1 degradation, and lysosomal acidification. It also modulates apoptosis by enhancing BAX/BAK-mediated mitochondrial outer membrane permeabilization. Mechanistically, DRAM1 interacts with LC3 family members, p62, and lysosomal proteins such as LAMP1 and cathepsins, linking p53 activation to both autophagic flux and cell death execution.
In the HAP1 background, DRAM1 disruption provides a clean genetic model to dissect autophagy and apoptosis crosstalk. Near-haploidy ensures that a single functional allele is targeted, leading to unambiguous loss-of-function phenotypes across the polyclonal population. Knockout cells display impaired autophagic flux, evidenced by accumulation of LC3-II and p62, and reduced lysosomal degradative capacity. This model allows researchers to directly assess how DRAM1 coordinates p53-dependent cellular responses without interference from wild-type alleles.
This knockout cell pool is highly applicable to autophagy research, cancer biology, neurodegenerative disease modeling, and infectious disease studies. Representative assays include western blotting for LC3 lipidation and p62 turnover, bafilomycin A1-based autophagy flux analysis, immunofluorescence colocalization of LAMP1 and LC3, Annexin V/PI apoptosis assays, qRT-PCR for autophagy-related genes, and lysosomal pH measurements. The model supports high-content screening and drug testing for autophagy modulators. For additional technical details or to discuss your specific experimental needs, please contact Ascent Research.
