SPP CRISPR Activation Plasmid (h): sc-405271-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SPP CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- SPP 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 SPP CRISPR Activation Plasmid (h) and SPP CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the HM13 transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SPP CRISPR Activation Plasmid (h) | sc-405271-ACT | 20 µg | $397.00 |
HM13 encodes signal peptide peptidase (SPP), an intramembrane aspartyl protease localized to the endoplasmic reticulum that cleaves remnant signal peptides after signal peptidase processing. By mediating regulated intramembrane proteolysis, SPP contributes to membrane protein turnover, peptide fragment generation, and coordination of ER protein quality control and secretory pathway homeostasis. SPP activity intersects with proteostasis programs such as ER-associated degradation and can influence antigenic peptide availability for immune surveillance through processing of signal peptide-derived substrates. Dysregulation of these pathways is relevant to cellular stress adaptation and immune-related mechanisms that are frequently perturbed in complex disease contexts.
SPP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HM13 expression without altering the underlying DNA sequence.
SPP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HM13 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 HM13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HM13 locus and enabling the study of SPP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPP pathway restoration in tumor cells with silenced or reduced HM13 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.