Ini1 CRISPR Activation Plasmid (h): sc-401485-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ini1 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Ini1 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 Ini1 CRISPR Activation Plasmid (h) and Ini1 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the SMARCB1 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: Ini1 Antibody (A-5): sc-166165
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ini1 CRISPR Activation Plasmid (h) | sc-401485-ACT | 20 µg | $397.00 | |||
Ini1 CRISPR Activation Plasmid (h2) | sc-401485-ACT-2 | 20 µg | $397.00 |
SMARCB1 (Ini1) encodes a core subunit of the SWI/SNF (BAF) ATP-dependent chromatin remodeling complex that regulates nucleosome positioning and chromatin accessibility. Through broad control of transcriptional programs, Ini1 influences cell-cycle progression, differentiation, lineage specification, and DNA damage responses by modulating enhancer and promoter activity. SMARCB1 perturbation is linked to disrupted epigenetic regulation and altered signaling networks, with strong relevance to tumor biology, including SMARCB1-deficient malignancies and other cancers featuring SWI/SNF dysfunction. These features make SMARCB1 a high-value target for studying chromatin-state transitions, transcription factor cooperativity, and genome-wide gene regulation.
Ini1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMARCB1 expression without altering the underlying DNA sequence.
Ini1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMARCB1 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 SMARCB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ini1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMARCB1 locus and enabling the study of Ini1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ini1 pathway restoration in tumor cells with silenced or reduced SMARCB1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.