Lyn CRISPR Activation Plasmid (h): sc-400463-ACT

Human LYN encodes Lyn, a Src family non-receptor tyrosine kinase that couples immunoreceptor and cytokine receptor signaling to downstream phosphorylation cascades. Lyn regulates B cell receptor and Fc receptor pathways, tuning activation thresholds through coordinated control of PI3K/AKT, MAPK, and NF-κB signaling and by modulating phosphatase recruitment. In hematopoietic cells it influences proliferation, survival, migration, and innate immune responses, while in other contexts it can affect adhesion and cytoskeletal dynamics via integrin-associated signaling. Dysregulated LYN activity or expression has been linked to altered immune homeostasis and signaling rewiring observed in hematologic malignancies and inflammatory disease mechanisms.

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

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

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