α-internexin CRISPR Activation Plasmid (h): sc-402920-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- α-internexin CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- α-internexin 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 α-internexin CRISPR Activation Plasmid (h) and α-internexin CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the INA 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: α-internexin Antibody (G-9): sc-271302
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
α-internexin CRISPR Activation Plasmid (h) | sc-402920-ACT | 20 µg | $397.00 | |||
α-internexin CRISPR Activation Plasmid (h2) | sc-402920-ACT-2 | 20 µg | $397.00 |
INA encodes α-internexin, a neuron-enriched type IV intermediate filament protein that integrates into the neurofilament network to support axonal caliber, cytoskeletal stability, and neurite outgrowth. α-internexin participates in intermediate filament assembly dynamics and coordinates with other neuronal cytoskeletal components during differentiation and synaptic maturation. Altered expression or aggregation of neuronal intermediate filaments is linked to mechanisms relevant to neurodegeneration and nervous system injury, including disrupted axonal transport and proteostasis stress. As a lineage-associated marker, INA is also used to study neuronal identity and differentiation programs in developmental and stem cell-derived neural models.
α-internexin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous INA expression without altering the underlying DNA sequence.
α-internexin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the INA 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 INA transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous α-internexin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native INA locus and enabling the study of α-internexin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of α-internexin pathway restoration in tumor cells with silenced or reduced INA expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.