Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
SPOP CRISPR Activation Plasmid (h): sc-401290-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SPOP CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- SPOP 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 SPOP CRISPR Activation Plasmid (h) and SPOP CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the SPOP 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: SPOP Antibody (B-8): sc-377206
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SPOP CRISPR Activation Plasmid (h) | sc-401290-ACT | 20 µg | $397.00 | |||
SPOP CRISPR Activation Plasmid (h2) | sc-401290-ACT-2 | 20 µg | $397.00 |
Human SPOP encodes speckle-type POZ protein, an adaptor for CUL3-based E3 ubiquitin ligase complexes that recognizes substrate proteins through its MATH domain and facilitates their ubiquitination and proteasomal turnover. By regulating the stability of key signaling and transcriptional regulators, SPOP influences nuclear protein homeostasis, DNA damage responses, and cell-cycle-associated programs. Altered SPOP function or expression has been linked to dysregulated proteostasis and transcriptional control in multiple disease contexts, supporting its use as a mechanistic node for studying ubiquitin-dependent signaling networks. These features make SPOP a useful target for probing pathway rewiring driven by changes in ubiquitin ligase substrate selection and turnover.
SPOP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPOP expression without altering the underlying DNA sequence.
SPOP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPOP 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 SPOP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SPOP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPOP locus and enabling the study of SPOP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SPOP pathway restoration in tumor cells with silenced or reduced SPOP expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.