JAK1 CRISPR Activation Plasmid (m): sc-421199-ACT

Mouse Jak1 encodes Janus kinase 1 (JAK1), a non-receptor tyrosine kinase that couples cytokine receptors to downstream STAT transcription factors to coordinate innate and adaptive immune signaling. JAK1 is central to interferon, IL-6 family, and common γ-chain cytokine pathways, shaping antiviral responses, inflammatory gene programs, and lymphocyte development. In addition to JAK–STAT, JAK1-driven signaling intersects with MAPK and PI3K/AKT networks to influence proliferation, survival, and differentiation. Dysregulated JAK1 activity or expression is frequently studied in inflammatory pathobiology and immune-associated disease mechanisms, as well as in oncogenic signaling contexts where cytokine cues remodel tumor–immune interactions.

JAK1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Jak1 expression without altering the underlying DNA sequence.

JAK1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Jak1 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 Jak1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous JAK1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Jak1 locus and enabling the study of JAK1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of JAK1 pathway restoration in tumor cells with silenced or reduced Jak1 expression.

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