PITSLRE A CRISPR Activation Plasmid (h): sc-402476-ACT

CDK11B encodes the human PITSLRE A serine/threonine kinase, a member of the CDC2L family implicated in cell-cycle–linked signaling and transcriptional control. PITSLRE A has been connected to RNA polymerase II–associated processes, RNA splicing and mRNA processing, and coordination of proliferation and stress responses through phosphorylation-dependent protein networks. As part of these regulatory circuits, altered CDK11B expression or activity is studied in contexts of dysregulated cell growth, genome maintenance, and apoptosis-related pathways that are frequently perturbed in disease biology. CDK11B is therefore a useful target for dissecting kinase-driven control of gene expression programs and cell-state transitions in human model systems.

PITSLRE A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDK11B expression without altering the underlying DNA sequence.

PITSLRE A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDK11B 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 CDK11B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PITSLRE A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDK11B locus and enabling the study of PITSLRE A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PITSLRE A pathway restoration in tumor cells with silenced or reduced CDK11B expression.

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