The ASRGL1 Knockout MCF-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from MCF-7 human breast adenocarcinoma cells, designed to disrupt the ASRGL1 gene. This polyclonal pool enables functional study of ASRGL1 in a luminal epithelial, estrogen receptor-positive breast cancer model. MCF-7 cells, isolated from a pleural effusion of a 69-year-old female with metastatic breast adenocarcinoma, retain ER?? and progesterone receptor expression, lack HER2 amplification, and serve as a standard model for hormonal therapy research.
MCF-7 cells exhibit a luminal epithelial phenotype and estrogen-dependent growth. As a TP53-wild-type, ER-positive model, they enable dissection of cross-talk between p53 and steroid hormone pathways. CRISPR-mediated disruption of ASRGL1 in this genetic context allows interrogation of its tumor-suppressive functions in a disease-relevant cellular environment.
ASRGL1 encodes an L-asparaginase and isoaspartyl peptidase that hydrolyzes L-asparagine to aspartic acid and ammonia, and removes isoaspartyl-damaged proteins. As a direct transcriptional target of TP53, ASRGL1 participates in a signaling network that includes CDKN1A, BAX, ASNS, and mTOR. Knockout elevates asparagine levels, activating mTORC1 and promoting proliferation. ASRGL1 also maintains protein homeostasis through isoaspartyl dipeptidase activity; its loss is predicted to impair apoptotic signaling due to accumulation of abnormal proteins.
In breast cancer, ASRGL1 is a putative tumor suppressor, with reduced expression in some tumors. This knockout in a TP53-wild-type, ER??-positive background creates a tool for studying p53-mediated metabolic tumor suppression. The model reveals how asparagine metabolism influences mTORC1-dependent proliferation in luminal breast cancer cells, where estrogen and amino acid signals converge. The polyclonal nature may reflect tumor heterogeneity, aiding preclinical evaluation of therapies targeting the p53-ASRGL1 axis.
Applications include mechanistic studies of p53 target genes, mTORC1 signaling regulation by asparagine, and drug response profiling for asparaginase-based therapies. Assays such as Western blotting for ASRGL1 and phospho-S6K1, proliferation (MTS/MTT), cell cycle flow cytometry, and Annexin V apoptosis assays are representative. Xenograft models can assess tumorigenicity and therapeutic responses. This polyclonal knockout cell population provides a robust platform for functional genomics and preclinical development in ER-positive breast cancer. For further details, contact Ascent Research.
