HLA-A CRISPR Activation Plasmid (h): sc-400294-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HLA-A CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- HLA-A CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by HLA-A CRISPR Activation Plasmid (h) and HLA-A CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the HLA-A transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HLA-A Antibody (B-11): sc-390473
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HLA-A CRISPR Activation Plasmid (h) | sc-400294-ACT | 20 µg | $397.00 |
HLA-A encodes a classical MHC class I heavy chain that pairs with β2-microglobulin to present endogenously derived peptides to CD8+ T cells, shaping immune surveillance and antigen-specific cytotoxic responses. Its expression and peptide repertoire integrate signals from the antigen processing and presentation pathway, including proteasomal processing, TAP-dependent peptide transport, and ER loading, and are modulated by interferon-driven JAK/STAT signaling. Variation in HLA-A influences allorecognition and immune compatibility and is widely studied in infection, tumor immunology, and autoimmune disease contexts where antigen presentation and immune evasion are key biological processes.
HLA-A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HLA-A expression without altering the underlying DNA sequence.
HLA-A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HLA-A locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.
Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the HLA-A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HLA-A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HLA-A locus and enabling the study of HLA-A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HLA-A pathway restoration in tumor cells with silenced or reduced HLA-A expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.