The DRAM2 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DRAM2 gene in human Huh-7 hepatocellular carcinoma cells. This model enables functional studies of DRAM2??s roles in autophagy and lysosomal regulation without the need for single-cell cloning. The polyclonal nature reflects heterogeneous gene disruption, providing a robust system for investigating autophagy, lysosomal function, and cell fate decisions in liver cancer research.
Huh-7 is a well-differentiated human hepatocellular carcinoma cell line established from a liver tumor of a 57-year-old male. Widely used for studying hepatocyte biology, hepatitis C virus infection, and liver cancer, Huh-7 cells retain malignant hepatocyte characteristics and key signaling pathways. Their compatibility with CRISPR/Cas9 editing allows precise gene disruption, making them an ideal host for examining DRAM2-dependent lysosome-mediated processes relevant to HCC progression.
DRAM2 encodes a lysosomal transmembrane protein regulated by TFEB, MITF, and mTORC1 inhibition. It promotes autophagic flux downstream of ULK1 and ATG5-ATG12, facilitating LC3-II processing and p62/SQSTM1 degradation. DRAM2 interacts with LAMP1, LAMP2, and cathepsins such as cathepsin B and cathepsin D, modulating lysosomal proteolysis. This protein integrates stress signals from DNA damage and nutrient deprivation to influence apoptosis through effectors like BAX and BCL-2, thereby connecting lysosomal function to cell death pathways.
In hepatocellular carcinoma, DRAM2-mediated autophagy and lysosomal degradation are vital for stress adaptation and therapy resistance. Knocking out DRAM2 in Huh-7 cells unveils how impaired lysosomal function affects tumor cell survival, drug sensitivity to agents like sorafenib, and innate immune signaling. This model is essential for dissecting crosstalk between mTORC1, lysosomal activity, and apoptosis, particularly given the role of autophagy in HCC progression and DRAM2??s link to p53-mediated DNA damage responses.
This knockout tool supports diverse applications such as autophagy regulation studies, lysosomal function analysis, and drug sensitivity testing using assays like Western blotting (LC3-II, p62), autophagy flux assays with chloroquine, immunofluorescence for LAMP1/2, apoptosis assays (Annexin V/PI), RT-qPCR, RNA-seq, and colony formation. It is also suitable for exploring innate immune pathways and identifying therapeutic targets in HCC. For further inquiries, please contact Ascent Research.
