JAK3 CRISPR Activation Plasmid (h): sc-400835-ACT

JAK3 (Janus kinase 3) is a non-receptor tyrosine kinase that associates with the common γ-chain (IL2RG) and transduces signals from multiple interleukin receptors, including IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21. Upon cytokine stimulation, JAK3 activates downstream STAT proteins and coordinates transcriptional programs that control lymphocyte development, activation, and survival, linking JAK–STAT signaling to immune homeostasis. Dysregulated JAK3 activity or expression perturbs cytokine signaling networks and has been implicated in primary immunodeficiency phenotypes as well as aberrant signaling observed in hematologic malignancies. Because JAK3 function is tightly coupled to immune cell fate decisions, it is widely studied in pathways governing T cell and NK cell biology and inflammatory signaling.

JAK3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous JAK3 expression without altering the underlying DNA sequence.

JAK3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the JAK3 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 JAK3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous JAK3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native JAK3 locus and enabling the study of JAK3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of JAK3 pathway restoration in tumor cells with silenced or reduced JAK3 expression.

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