Human BAK shRNA Lentivirus

Background

BAK, encoded by BAK1, is a pro-apoptotic effector of the BCL-2 protein family and, together with BAX, executes the intrinsic apoptotic pathway. Following apoptotic signals, activated BAK oligomerizes within the mitochondrial outer membrane and forms pores that permeabilize the membrane, releasing cytochrome c and other factors that trigger caspase activation and cell death. BAK activity is restrained by anti-apoptotic BCL-2 proteins and promoted by BH3-only proteins, placing it at the convergence point of survival and death signals. Dysregulation of BAK-dependent apoptosis contributes to tumorigenesis and resistance to chemotherapy, making it a key target in cancer and cell death research.

Product Description & Applications

The Human BAK shRNA Lentivirus delivers validated short hairpin RNA targeting BAK1 from a third-generation, self-inactivating lentiviral backbone. shRNA expression is driven by a U6 promoter, with a co-expressed fluorescent reporter (GFP or RFP) and antibiotic selection marker. VSV-G pseudotyping enables broad tropism across primary, suspension, and cryopreserved cells, and the shRNA is validated for at least 70% BAK knockdown by a fluorescence-based method.

High-titer particles are ultra-purified by PEG precipitation and sucrose gradient centrifugation, suitable for transducing difficult-to-transfect cells. Applications include loss-of-function studies of mitochondrial apoptosis, investigations of cell death regulation, and analysis of chemotherapy resistance mechanisms.

About This Product

This validated shRNA lentivirus targeting BAK1 (NCBI Accession: NM_001188.4) 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.