Src CRISPR Activation Plasmid (h): sc-400165-ACT

Human SRC encodes Src, a prototypical non-receptor tyrosine kinase that integrates signals from receptor tyrosine kinases, integrins, and G protein–coupled receptors to coordinate proliferation, survival, adhesion, and cytoskeletal remodeling. Src activity propagates through pathways including focal adhesion signaling, PI3K–AKT, RAS–MAPK, and STAT to modulate cell motility and transcriptional programs. Dysregulated SRC signaling is frequently associated with invasive behavior, altered cell–matrix interactions, and aberrant growth factor responsiveness across diverse cancer models. SRC also contributes to immune receptor signaling and endothelial barrier dynamics, supporting its broad relevance in systems biology and disease mechanism research.

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

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

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