Sprouty 2 CRISPR Activation Plasmid (h): sc-401087-ACT

SPRY2 encodes Sprouty 2, a conserved feedback inhibitor of receptor tyrosine kinase signaling that modulates growth factor–driven outputs such as proliferation, differentiation, and migration. Sprouty 2 primarily dampens RAS–RAF–MEK–ERK/MAPK signaling downstream of FGFR, EGFR, and other RTKs by influencing adaptor and effector interactions, thereby shaping signal intensity and duration. Through these functions, SPRY2 contributes to regulation of developmental programs, epithelial–mesenchymal dynamics, and cellular stress responses. Altered SPRY2 expression or regulation has been associated with dysregulated MAPK pathway activity observed in multiple disease-relevant contexts, including oncogenic signaling networks and aberrant tissue remodeling.

Sprouty 2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SPRY2 expression without altering the underlying DNA sequence.

Sprouty 2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SPRY2 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 SPRY2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Sprouty 2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SPRY2 locus and enabling the study of Sprouty 2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Sprouty 2 pathway restoration in tumor cells with silenced or reduced SPRY2 expression.

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