Human PDL1 shRNA Lentivirus

Background

CD274, also known as PD-L1 or B7-H1, is a cell-surface ligand that delivers an inhibitory signal to T cells through the PD-1 receptor. Engagement of PD-1 by PD-L1 suppresses T-cell receptor signaling, proliferation, and cytokine production, contributing to peripheral tolerance and the prevention of excessive immune responses. Many tumors and antigen-presenting cells upregulate PD-L1 in response to inflammatory signals such as interferon-gamma, exploiting this checkpoint to evade T-cell-mediated killing. The PD-1/PD-L1 axis is a central target of cancer immunotherapy, and PD-L1 expression is widely studied as a determinant of tumor immune evasion and as a biomarker for checkpoint-blockade therapy.

Product Description & Applications

The Human PDL1 shRNA Lentivirus delivers validated short hairpin RNA targeting human CD274 (PD-L1) for stable gene knockdown. Each shRNA is validated to achieve at least 70% reduction of PD-L1 using a fluorescence-based assay that provides a direct functional readout rather than transcript-level qPCR. The particles are ultra-purified and concentrated to high titer by PEG precipitation and sucrose gradient centrifugation, and efficiently transduce difficult-to-transfect cells, including primary and thawed cultures. A co-expressed fluorescent reporter (GFP or RFP, optionally with luciferase) and antibiotic selection marker (puromycin or blasticidin) enable stable line generation. A shRNA set option supplies a mix of two independent validated shRNAs plus matched scrambled-control lentivirus for loss-of-function studies of immune checkpoint signaling and tumor immune evasion.

About This Product

This validated shRNA lentivirus targeting CD274 (NCBI Accession: NM_014143) delivers a 19–20 bp shRNA from a third-generation, self-inactivating lentiviral backbone. Expression is driven from a U6 Pol III promoter, with a constitutively expressed fluorescent reporter (GFP, GFP/Luc, RFP, RFP/Luc) and antibiotic selection marker (Blasticidin, Puromycin) co-expressed from the same vector. VSV-G pseudotyping enables broad cell tropism, including primary, suspension, and cryopreserved cell types.

Knockdown is validated using a proprietary bicistronic fluorescence assay in which the target mRNA is co-expressed fused to RFP alongside the shRNA-GFP construct. At least 70% reduction in RFP signal in GFP-positive cells confirms on-target activity — a more direct functional readout than transcript-level qPCR. Polyclonal stable lines can be generated by antibiotic selection within 10 days, preserving parental cell heterogeneity compared to single-clone CRISPR approaches.