| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Product Type | |
| Promoter | |
| Reporter | |
| Selection Marker | Blasticidin, N/A, Puromycin |
| Shipping | |
| Species |
Background
H3.3, encoded by H3-3A, is a replication-independent variant of histone H3 that is incorporated into nucleosomes throughout the cell cycle, particularly at actively transcribed genes, enhancers, and telomeric and pericentric heterochromatin. By marking dynamic chromatin, H3.3 contributes to transcriptional regulation, genome stability, and epigenetic inheritance. Recurrent oncohistone mutations in H3.3, such as K27M and G34 substitutions, are key drivers of pediatric high-grade gliomas and certain bone and cartilage tumors, where they reprogram the epigenome. Wild-type and mutant H3.3 are therefore widely studied in chromatin biology and cancer research.
Product Description & Applications
The H3.3 (WT & Mutant) ORF cDNA Lentivirus delivers the human H3.3 (H3-3A) open reading frame for stable overexpression in mammalian cells, available in wild-type and mutant forms for comparative studies. Expression of the ORF, fused to a C-terminal V5 tag, is driven by an EF1α or CMV promoter, with a GFP or RFP reporter and optional blasticidin or puromycin selection marker separated by self-cleaving peptide sequences for independent translation. The lentiviral vectors are optimized for high expression and reliable genome integration; ORF accuracy is confirmed by sequencing and protein expression validated by transient transfection.
Supplied as high-titer, third-generation, VSV-G-pseudotyped particles, it efficiently transduces primary and thawed cells for studying H3.3 function and oncohistone mutations.
About This Product
This ORF cDNA lentivirus enables stable overexpression of H3-3A (NCBI Accession: NM_002107.7) in mammalian cells via a third-generation, VSV-G pseudotyped delivery system. The ORF cDNA is fused to a C-terminal epitope tag (V5, Myc, or HA) and expressed under a strong constitutive promoter (EF1a). Reporter and selection marker components (GFP, RFP; Blasticidin, Puromycin) are co-expressed via self-cleaving P2A peptides, enabling independent protein production without fusion-tag artifacts.
Ultra-purification by PEG precipitation and sucrose gradient centrifugation yields high-titer particles suitable for primary cells, suspension cultures, and stem cells. Stable polyclonal cell lines are established within 10–14 days by antibiotic selection or FACS sorting. For in vivo applications, the serum-free formulation and VSV-G envelope support direct administration or further concentration for stereotactic injection.
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.
