Pim-2 CRISPR Activation Plasmid (h): sc-401715-ACT

PIM2 encodes Pim-2, a constitutively active serine/threonine kinase that integrates cytokine and growth factor inputs to promote cell survival, proliferation, and metabolic adaptation. Pim-2 phosphorylates signaling and apoptotic regulators, supporting translational control and resistance to cellular stress, and functionally intersects with JAK/STAT, PI3K/AKT/mTOR, and NF-κB-associated programs. Dysregulated PIM2 expression or activity has been linked to altered hematopoietic cell homeostasis and oncogenic signaling networks, particularly in contexts where survival pathways are persistently engaged. As a modulator of phosphorylation-dependent checkpoints, Pim-2 is frequently studied for its effects on apoptosis, cell-cycle progression, and therapy response phenotypes in model systems.

Pim-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIM2 expression without altering the underlying DNA sequence.

Pim-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIM2 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 PIM2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pim-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIM2 locus and enabling the study of Pim-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pim-2 pathway restoration in tumor cells with silenced or reduced PIM2 expression.

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