L-type Ca++ CP α1S CRISPR Activation Plasmid (h): sc-402713-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- L-type Ca++ CP α1S CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- L-type Ca++ CP α1S 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 L-type Ca++ CP α1S CRISPR Activation Plasmid (h) and L-type Ca++ CP α1S CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the CACNA1S 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: L-type Ca++ CP α1S Antibody (G-1): sc-514685
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
L-type Ca++ CP α1S CRISPR Activation Plasmid (h) | sc-402713-ACT | 20 µg | $397.00 |
CACNA1S encodes the α1S pore-forming subunit of the skeletal muscle L-type voltage-gated calcium channel (CaV1.1), a key component of the dihydropyridine receptor complex in the transverse tubule membrane. Upon membrane depolarization, CaV1.1 couples electrically to ryanodine receptor 1 to initiate excitation–contraction coupling and regulate calcium flux that shapes myofiber contraction dynamics. This channel participates in calcium-dependent signaling networks that influence muscle development, metabolic homeostasis, and activity-dependent gene expression. Genetic and functional perturbations in CACNA1S have been associated with skeletal muscle channelopathies and susceptibility to dysregulated calcium handling, supporting its relevance in studies of neuromuscular physiology and disease mechanisms.
L-type Ca++ CP α1S CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CACNA1S expression without altering the underlying DNA sequence.
L-type Ca++ CP α1S CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CACNA1S 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 CACNA1S transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous L-type Ca++ CP α1S expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CACNA1S locus and enabling the study of L-type Ca++ CP α1S-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of L-type Ca++ CP α1S pathway restoration in tumor cells with silenced or reduced CACNA1S expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.