The GPR171 Knockout MCF-7 Polyclonal Cells are a CRISPR/Cas9-edited human polyclonal cell population derived from the MCF-7 breast adenocarcinoma line. This model harbors targeted disruptions of the GPR171 gene introduced via CRISPR/Cas9, generating a heterogeneous pool of cells carrying loss-of-function mutations at the GPR171 locus. Supplied as a polyclonal knockout population, it avoids clonal selection biases and facilitates functional analysis in a bulk population context, capturing diverse cellular responses to gene inactivation.
The parental MCF-7 cell line is a widely characterized model originally isolated from a pleural effusion of a metastatic mammary adenocarcinoma. These cells are estrogen receptor (ER)-positive and progesterone receptor (PR)-positive, retaining key features of hormone-responsive breast cancer. MCF-7 has been pivotal in elucidating estrogen signaling mechanisms, endocrine therapy resistance, and tumor progression, and serves as a cornerstone for oncology research, particularly for studies of the luminal A breast cancer subtype.
GPR171 encodes a G protein-coupled receptor that functions as a high-affinity receptor for the endogenous neuropeptide BigLEN, a product of the PCSK1N gene. Upon ligand binding, GPR171 couples to G??i/o proteins, leading to inhibition of adenylate cyclase, decreased intracellular cAMP, and suppression of protein kinase A (PKA). Through interactions with ??-arrestin-1 and ??-arrestin-2, the receptor triggers the RAS-RAF-MEK-ERK MAPK cascade and the PI3K-AKT pathway, culminating in transcriptional activation of FOS, JUN, and MYC. In MCF-7 cells, GPR171 activity is also influenced by estrogen signaling, establishing crosstalk between hormone and neuropeptide pathways.
Within the MCF-7 background, GPR171 sits at the nexus of GPCR, metabolic, and hormone-dependent oncogenic signaling. Disruption of the GPR171 gene is expected to attenuate BigLEN-driven MAPK/ERK and PI3K/AKT activation, thereby diminishing the proliferative and survival signals that contribute to aggressive tumor phenotypes. This knockout model allows precise dissection of G protein-coupled receptor contributions to estrogen-responsive breast cancer progression and enables investigation of whether GPR171 signaling intersects with classical estrogen receptor genomic and non-genomic actions.
This polyclonal knockout population supports a range of downstream applications, including western blotting, RT-qPCR, and RNA-seq to assess molecular changes, as well as cell proliferation, migration, and drug sensitivity assays for functional phenotyping. Quantitative signaling readouts can be obtained via cAMP ELISA and phospho-ERK/AKT flow cytometry. These approaches facilitate target validation, mechanistic studies of neuropeptide-driven oncogenic pathways, and preclinical evaluation of therapeutic strategies for breast cancer and metabolic disorders. For further technical details, please contact Ascent Research.
