Yes CRISPR Activation Plasmid (m): sc-423744-ACT

Mouse Yes1 encodes a non-receptor tyrosine kinase of the SRC family that transduces signals downstream of receptor tyrosine kinases and cell–cell adhesion complexes to regulate proliferation, survival, and cytoskeletal remodeling. YES1 activity interfaces with MAPK/ERK and PI3K/AKT signaling and can influence focal adhesion dynamics and cell motility through phosphorylation of adaptor and scaffold proteins. Altered YES1 signaling has been linked to oncogenic signaling states, drug response phenotypes, and rewiring of growth-factor–driven pathways in multiple tumor contexts. In basic and translational research models, Yes1 modulation supports studies of signaling network redundancy among SRC family kinases and pathway cross-talk controlling differentiation and tissue homeostasis.

Yes CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Yes1 expression without altering the underlying DNA sequence.

Yes CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Yes1 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 Yes1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Yes expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Yes1 locus and enabling the study of Yes-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Yes pathway restoration in tumor cells with silenced or reduced Yes1 expression.

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