c-Kit CRISPR Activation Plasmid (m): sc-421289-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- c-Kit CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- c-Kit CRISPR Activation Plasmid (m) 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 c-Kit CRISPR Activation Plasmid (m) and c-Kit CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Kit 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: c-Kit Antibody (E-3): sc-365504
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
c-Kit CRISPR Activation Plasmid (m) | sc-421289-ACT | 20 µg | $397.00 |
Mouse Kit encodes c-Kit (CD117), a type III receptor tyrosine kinase that binds stem cell factor to regulate hematopoietic stem and progenitor maintenance, melanocyte development, and germ cell survival. Upon ligand-induced dimerization and autophosphorylation, c-Kit activates PI3K–AKT, RAS–MAPK, JAK–STAT, and PLCγ signaling to control proliferation, migration, and differentiation. Dysregulated KIT signaling and altered c-Kit expression are implicated in aberrant mast cell biology and hematologic malignancy models, and are widely studied in developmental biology and immunology. As a surface receptor with well-defined downstream pathways, c-Kit serves as a tractable node for probing signaling thresholds and lineage decisions in mouse systems.
c-Kit CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Kit expression without altering the underlying DNA sequence.
c-Kit CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Kit 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 Kit transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous c-Kit expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Kit locus and enabling the study of c-Kit-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of c-Kit pathway restoration in tumor cells with silenced or reduced Kit expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.