MACC1 CRISPR Activation Plasmid (h): sc-404253-ACT

Human MACC1 (metastasis associated in colon cancer 1) encodes a transcriptional regulator implicated in control of cell motility, invasive growth, and survival programs. MACC1 is closely linked to HGF/MET signaling and downstream PI3K–AKT and MAPK pathway activity, coordinating gene expression changes that support epithelial–mesenchymal transition–like phenotypes and remodeling of cytoskeletal and adhesion dynamics. Altered MACC1 expression has been associated with tumor progression and metastatic behavior across multiple solid malignancies, making it a useful molecular node for studying transcriptional networks that couple growth factor sensing to migration. Its regulation and downstream targets are therefore frequently interrogated in cancer biology models, including studies of invasion, metastasis-associated transcriptional signatures, and pathway cross-talk.

MACC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MACC1 expression without altering the underlying DNA sequence.

MACC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MACC1 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 MACC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MACC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MACC1 locus and enabling the study of MACC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MACC1 pathway restoration in tumor cells with silenced or reduced MACC1 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.