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| Selection Marker | Blasticidin, N/A, Puromycin |
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Background
PUMA (p53 upregulated modulator of apoptosis), encoded by BBC3, is a BH3-only member of the BCL-2 protein family and a key effector of apoptosis. PUMA expression is induced by p53 following DNA damage and by p53-independent stimuli such as growth factor withdrawal and cellular stress. Once expressed, PUMA binds and inhibits anti-apoptotic BCL-2 proteins, freeing BAX and BAK to permeabilize the mitochondrial outer membrane and trigger caspase activation. Through this role, PUMA links diverse stress signals to programmed cell death and contributes to tumor suppression, making it an important target in cancer research, apoptosis studies, and investigations of treatment resistance.
Product Description & Applications
The Human PUMA shRNA Lentivirus delivers validated short hairpin RNA targeting BBC3 from a third-generation, self-inactivating lentiviral backbone, with shRNA expression driven by a U6 promoter and a co-expressed fluorescent reporter (GFP or RFP) and antibiotic selection marker. VSV-G pseudotyping supports broad tropism, including primary, suspension, and cryopreserved cells. Each shRNA is validated to achieve at least 70% PUMA knockdown using a fluorescence-based assay.
The high-titer particles are ultra-purified by PEG precipitation and sucrose gradient centrifugation. A shRNA set option provides a mix of two independent validated shRNAs plus a scrambled control. Applications include loss-of-function studies of apoptosis, tumor suppression, and stress responses, and generation of stable knockdown cell lines.
About This Product
This validated shRNA lentivirus targeting BBC3 (NCBI Accession: NM_014417.5) 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, RFP, GFP/Luc, 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.
Can't find the lentiviral construct you need, or want to adjust key design elements? Contact us to discuss custom LV design and optional add-ons.
Common customization requests
- Insert / payload: replace the gene/sequence, swap to a different isoform, add mutations, or optimize cloning features.
- Expression design: change promoter (e.g., CMV/EF1α/PGK), add enhancers, or adjust regulatory elements.
- Reporters: add/swap GFP/RFP/mCherry/luciferase (single or dual reporters where applicable).
- Selection markers: add/swap puromycin/blasticidin/neomycin or fluorescent selection options.
- Vector format: switch between OE, shRNA, CRISPR (sgRNA/Cas systems), or control vectors (where supported).
Add-ons you can request
- Control viruses: empty vector, non-targeting shRNA, reporter-only controls, or matched backbone controls.
- Packaging / format: concentration options, aliquoting, or custom fill volume for screening workflows.
- Documentation: construct map/sequence confirmation package (as available) and batch documentation.
What to include in your request
- Target cell type/model (cell line or primary cells) and intended readout (reporter, knockdown, OE, etc.)
- Insert sequence (FASTA) or reference ID, plus any required tags/mutations
- Promoter, reporter, and selection marker preferences
- Desired scale and preferred format (aliquots / concentration requests)
Email us at support@biohippo.com or use the Talk to a Scientist request form.
