Sp8 CRISPR Activation Plasmid (h): sc-401546-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Sp8 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Sp8 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 Sp8 CRISPR Activation Plasmid (h) and Sp8 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the SP8 transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Sp8 CRISPR Activation Plasmid (h) | sc-401546-ACT | 20 µg | $397.00 |
SP8 encodes a Sp8 zinc finger transcription factor that binds GC-rich regulatory elements to control gene expression programs during human embryonic development. It is implicated in patterning and differentiation processes, including neuroectodermal and craniofacial development, by coordinating transcriptional networks that interface with morphogen-driven signaling such as WNT/β-catenin and related developmental pathways. Altered SP8 activity or dysregulated target gene networks has been associated with congenital malformation phenotypes and aberrant developmental gene regulation, making it relevant for studies of lineage specification and morphogenesis. As a nuclear DNA-binding regulator, Sp8 is frequently used as a node for mapping transcriptional circuits and enhancer-promoter control in differentiating systems.
Sp8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SP8 expression without altering the underlying DNA sequence.
Sp8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SP8 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 SP8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sp8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SP8 locus and enabling the study of Sp8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sp8 pathway restoration in tumor cells with silenced or reduced SP8 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.