Scrib CRISPR Activation Plasmid (h): sc-400384-ACT

Human SCRIB encodes Scrib, a conserved polarity scaffold that localizes to basolateral membranes and coordinates assembly of protein complexes controlling apical–basal polarity, adherens junction integrity, and directed cell migration. Scrib interfaces with core polarity modules and signaling networks including Hippo pathway regulation of YAP/TAZ, MAPK signaling dynamics, and planar cell polarity processes that shape tissue architecture. Altered SCRIB expression or mislocalization is associated with disrupted epithelial organization, aberrant cytoskeletal remodeling, and changes in contact inhibition, phenotypes frequently observed in malignant transformation and metastatic progression. As a result, SCRIB is widely studied in models of epithelial homeostasis, invasion, and signal integration at cell–cell junctions.

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

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

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