cyclin T1 CRISPR Activation Plasmid (h): sc-400623-ACT

CCNT1 encodes cyclin T1, the regulatory partner of CDK9 in the positive transcription elongation factor b (P-TEFb) complex that stimulates RNA polymerase II pause release by phosphorylating the CTD and negative elongation factors. Through control of transcriptional elongation, cyclin T1 influences cell-cycle progression, DNA damage responses, and differentiation programs, and it is a central host factor co-opted by HIV-1 Tat to enhance viral transcription. Altered P-TEFb activity and cyclin T1 availability have been linked to dysregulated transcriptional networks observed in cancer biology and inflammatory signaling contexts. CCNT1 is therefore widely studied in pathways governing transcriptional control, chromatin-associated regulation, and stress-responsive gene expression.

cyclin T1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CCNT1 expression without altering the underlying DNA sequence.

cyclin T1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CCNT1 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 CCNT1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous cyclin T1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CCNT1 locus and enabling the study of cyclin T1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of cyclin T1 pathway restoration in tumor cells with silenced or reduced CCNT1 expression.

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