STRAD CRISPR Activation Plasmid (h): sc-402311-ACT

STRADA encodes STRAD, a catalytically inactive pseudokinase that forms a complex with LKB1 (STK11) and the scaffold protein MO25 to promote LKB1 cytoplasmic localization and activation. Through this complex, STRAD supports phosphorylation cascades that influence AMPK-related kinase signaling, cellular energy sensing, polarity, and growth control, with downstream impacts on processes such as cytoskeletal organization and stress responses. Disruption or altered regulation of STRADA has been linked to neurodevelopmental phenotypes and dysregulated signaling networks that intersect with pathways frequently studied in proliferation and migration biology. As a result, STRAD is commonly investigated as a regulatory node connecting kinase scaffolding, subcellular localization, and context-dependent transcriptional programs.

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

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

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