Stat5b CRISPR Activation Plasmid (h): sc-400878-ACT

Human STAT5B encodes Stat5b, a signal-dependent transcription factor activated downstream of cytokine and growth factor receptors via JAK kinases, with phosphorylation-driven dimerization and nuclear translocation. Stat5b regulates transcriptional programs controlling lymphocyte development, immune homeostasis, and somatic growth, and it integrates inputs from pathways including JAK–STAT and cross-talk with MAPK and PI3K signaling. Dysregulated STAT5B activity is associated with altered cytokine responsiveness and aberrant cell-state transitions relevant to immunologic disorders and hematologic malignancy biology. As a transcriptional hub, STAT5B is widely studied for its effects on proliferation, differentiation, survival, and lineage-specific gene expression.

Stat5b CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous STAT5B expression without altering the underlying DNA sequence.

Stat5b CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the STAT5B 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 STAT5B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Stat5b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native STAT5B locus and enabling the study of Stat5b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Stat5b pathway restoration in tumor cells with silenced or reduced STAT5B expression.

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