Pim-1 CRISPR Activation Plasmid (h): sc-400376-ACT

PIM1 encodes Pim-1, a constitutively active serine/threonine kinase that integrates cytokine and growth factor cues to regulate cell survival, proliferation, and metabolic fitness. Pim-1 modulates key signaling networks including JAK/STAT, PI3K/AKT, and NF-κB, and phosphorylates substrates that influence cell-cycle progression and apoptosis such as BAD, p21, and transcriptional regulators. Through these activities, PIM1 impacts hematopoietic and immune cell function and contributes to stress-adaptive programs that can be co-opted in oncogenic contexts. Dysregulated PIM1 expression and signaling have been associated with altered proliferative control, therapy resistance phenotypes in model systems, and disease-relevant transcriptional states in multiple cancer types.

Pim-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PIM1 expression without altering the underlying DNA sequence.

Pim-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PIM1 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 PIM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Pim-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PIM1 locus and enabling the study of Pim-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Pim-1 pathway restoration in tumor cells with silenced or reduced PIM1 expression.

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