MARCKS CRISPR Activation Plasmid (h): sc-400837-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- MARCKS CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- MARCKS 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 MARCKS CRISPR Activation Plasmid (h) and MARCKS CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the MARCKS 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: MARCKS Antibody (JK-8): sc-100777
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
MARCKS CRISPR Activation Plasmid (h) | sc-400837-ACT | 20 µg | $397.00 |
Myristoylated alanine-rich C kinase substrate (MARCKS) is a prominent PKC substrate that associates with the plasma membrane and actin cytoskeleton to regulate cell shape, adhesion, and motility. By binding phosphatidylinositol 4,5-bisphosphate (PIP2) and crosslinking F-actin, MARCKS influences membrane trafficking, exocytosis/endocytosis, and signal transduction downstream of PKC and Ca2+/calmodulin. MARCKS-dependent cytoskeletal remodeling is implicated in neurite outgrowth and synaptic plasticity as well as immune cell migration and inflammatory signaling. Dysregulated MARCKS expression or phosphorylation has been reported in cancer biology, neuroinflammation, and pulmonary disease contexts, supporting its use as a mechanistic node in studies of migration, invasion, and stimulus-dependent signaling.
MARCKS CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MARCKS expression without altering the underlying DNA sequence.
MARCKS CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MARCKS 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 MARCKS transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MARCKS expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MARCKS locus and enabling the study of MARCKS-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MARCKS pathway restoration in tumor cells with silenced or reduced MARCKS expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.