Cdk9 CRISPR Activation Plasmid (h): sc-400242-ACT

Human CDK9 encodes cyclin-dependent kinase 9 (Cdk9), the catalytic subunit of positive transcription elongation factor b (P-TEFb) that phosphorylates the RNA polymerase II C-terminal domain to promote productive transcriptional elongation. Through regulation of promoter-proximal pause release, Cdk9 coordinates expression of immediate-early genes, stress response programs, and cell-cycle and survival regulators, and integrates signals from transcriptional coactivators and chromatin-associated factors. CDK9 activity influences RNA processing and genome stability by shaping transcription-coupled events and controlling the kinetics of gene expression. Dysregulated CDK9/P-TEFb signaling is frequently associated with aberrant transcriptional dependencies relevant to cancer biology, inflammatory signaling, and virus–host transcriptional control, making it a widely used node for mechanistic studies of transcription.

Cdk9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDK9 expression without altering the underlying DNA sequence.

Cdk9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDK9 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 CDK9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cdk9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDK9 locus and enabling the study of Cdk9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cdk9 pathway restoration in tumor cells with silenced or reduced CDK9 expression.

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