This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the SK-HEP-1 human liver adenocarcinoma cell line, engineered to disrupt the gene encoding L3MBTL2 (Lethal(3)malignant brain tumor-like protein 2). L3MBTL2 functions as a chromatin regulator that recognizes mono- and dimethylated histone marks, specifically H4K20me1/2 and H3K9me1/2, to mediate transcriptional repression. The polyclonal knockout format provides a heterogeneous pool of cells harboring diverse loss-of-function mutations, reducing clonal bias and enabling robust functional genomics studies. This model is suitable for investigating the role of L3MBTL2 in DNA damage signaling, epigenetic silencing, and hepatocellular carcinoma progression.
The host cell line, SK-HEP-1, is a widely used human hepatocellular carcinoma model derived from ascitic fluid of a patient with liver adenocarcinoma. This cell line exhibits a mixed epithelial and endothelial phenotype, making it valuable for studying hepatic cancer biology, metastasis, and tumor microenvironment interactions. SK-HEP-1 cells display rapid proliferation and are readily amenable to transfection and lentiviral transduction, facilitating efficient CRISPR/Cas9-mediated gene disruption. The knockout cells are generated and maintained under standard culture conditions, ensuring compatibility with a broad range of downstream assays.
L3MBTL2 is a critical component of the DNA damage response and chromatin organization pathways. Upon DNA damage, L3MBTL2 binds methylated histones at break sites and interacts with TP53BP1 to facilitate non-homologous end joining and local transcriptional silencing. Its activity is regulated by upstream factors such as ATM kinase and histone methylation marks. L3MBTL2 forms complexes with PHF20 and KDM1A, and cooperates with its paralog L3MBTL1. Downstream, L3MBTL2 influences DNA repair gene expression, cell cycle regulators, and TP53 signaling components. Knockout of L3MBTL2 disrupts recruitment of repair factors, leading to defective genome stability maintenance and altered transcriptional programs.
In the SK-HEP-1 hepatocellular carcinoma context, loss of L3MBTL2 is expected to impair DNA repair fidelity and alter chromatin states, potentially affecting tumor-suppressive pathways. This polyclonal knockout model enables dissection of L3MBTL2??s role in liver cancer cell biology, including responses to genotoxic stress such as ionizing radiation or chemotherapeutic agents. The heterogeneity of the polyclonal population mimics the genetic variability seen in tumors, providing a more physiologically relevant system for studying tumorigenesis mechanisms and drug resistance phenotypes.
This knockout cell pool is ideal for a variety of experimental applications, including chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) to assess histone modification occupancy, western blotting and RT-qPCR for expression analysis, and RNA sequencing for transcriptome-wide effect characterization. Functional assays such as immunofluorescence microscopy for ??-H2AX foci formation, flow cytometric cell cycle profiling, and proliferation assays can be employed to evaluate DNA damage response and growth defects. Phospho-signaling analysis by immunoblotting or multiplex assays further enables exploration of ATM/TP53BP1 axis activation. For additional details or custom options, please contact Ascent Research.
