Genome-edited Cells
The CD274 Knockout U-87MG Cell Line is a CRISPR/Cas9-edited knockout cell line derived from human U-87MG glioblastoma cells. It features targeted disruption of CD274, encoding PD-L1, an immune checkpoint ligand that suppresses T cell activity via PD-1 engagement and downstream SHP2-mediated inhibition of PI3K/AKT and MAPK pathways. This model is ideal for studying PD-L1-dependent immune evasion in glioblastoma and for evaluating anti-PD-L1 therapeutics. Key applications include co-culture immune assays, cytokine profiling, and drug screening, leveraging techniques such as flow cytometry, western blot, and T cell cytotoxicity assays.
GLOD4 Knockout SK-HEP-1 Polyclonal Cells
Cat. No. ARG15370
NRM Knockout HT29 Polyclonal Cells
Cat. No. ARG14466
LSM1 Knockout HT29 Polyclonal Cells
Cat. No. ARG14026
KALRN Knockout HAP1 Polyclonal Cells
Cat. No. ARG22899
METTL13 Knockout 786-O Polyclonal Cells
Cat. No. ARG5749
Rabbit Renal Artery Smooth Muscle Cells
Cat. No. ARP0702
The CD274 Knockout U-87MG Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the human U-87MG glioblastoma cell line. This cell line features targeted disruption of the CD274 gene, which encodes the PD-L1 (B7-H1) immune checkpoint ligand, and serves as a validated loss-of-function model for interrogating PD-L1-mediated immune evasion mechanisms.
The parental U-87MG cell line is a well-characterized human glioblastoma model derived from a malignant glioma. It harbors a PTEN mutation, wild-type p53, and amplifies EGFR, making it a relevant system for studying signaling networks that drive glioblastoma multiforme. U-87MG cells are commonly employed to investigate tumor proliferation, invasion, and immune escape mechanisms.
The CD274 gene encodes programmed death-ligand 1 (PD-L1, B7-H1), a transmembrane immune checkpoint protein. PD-L1 interacts with its receptor PD-1 (PDCD1) on activated T cells, recruiting the tyrosine phosphatases SHP-1 (PTPN6) and SHP-2 (PTPN11) to the intracellular domain. This initiates an inhibitory cascade that dephosphorylates key TCR-proximal kinases such as ZAP70 and LCK, impairing downstream PI3K/AKT and RAS/MEK/ERK signaling, ultimately reducing IL-2 and IFN?? production and promoting T cell exhaustion. PD-L1 expression is transcriptionally regulated by IFN-?? through STAT1 and NF-??B, and by oncogenic signals including EGFR, PI3K/AKT/mTOR, and HIF-1??.
In U-87MG glioblastoma cells, PD-L1 is often upregulated as a mechanism of immune evasion, dampening anti-tumor T cell responses within the tumor microenvironment. Disruption of CD274 in this cell line abrogates PD-L1 expression, removing the inhibitory signal and potentially restoring T cell effector functions. This isogenic knockout model therefore provides a powerful tool to dissect the contribution of PD-L1 to glioblastoma immune escape and to assess the efficacy of PD-1/PD-L1 blockade strategies in a defined genetic background.
Researchers can employ this CD274 knockout cell line for a variety of immune-oncology applications, including co-culture assays with primary T cells or PBMCs to measure restoration of cytotoxicity and cytokine secretion (e.g., IFN??, IL-2 via ELISA), screening of novel anti-PD-L1 small molecules or biologics, and dissecting PD-L1-driven signaling crosstalk with EGFR, PI3K/AKT, and MAPK pathways. The loss-of-function model is particularly suited for comparative studies with the parental U-87MG line using techniques such as flow cytometry (to confirm PD-L1 ablation), western blotting, RT-qPCR, and immunofluorescence. It also enables investigation of tumor-intrinsic roles of PD-L1 in proliferation, migration, and drug sensitivity. For more information, please contact Ascent Research.
