Cdk7 CRISPR Activation Plasmid (h): sc-400665-ACT

Human CDK7 encodes cyclin-dependent kinase 7 (Cdk7), a dual-function enzyme that serves as the CDK-activating kinase within the CDK7–cyclin H–MAT1 complex and as a core component of general transcription factor TFIIH. By phosphorylating the RNA polymerase II C-terminal domain and activating cell-cycle CDKs, Cdk7 coordinates transcription initiation, promoter-proximal pause release, and orderly cell-cycle progression. CDK7 activity is tightly integrated with DNA damage response signaling and transcription-coupled nucleotide excision repair, linking genome maintenance to cell-fate decisions. Dysregulation of CDK7-dependent transcriptional programs and cell-cycle control is frequently studied in the context of proliferative states and oncogenic transcription networks, making CDK7 a high-value node for mechanistic pathway interrogation.

Cdk7 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CDK7 expression without altering the underlying DNA sequence.

Cdk7 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CDK7 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 CDK7 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Cdk7 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CDK7 locus and enabling the study of Cdk7-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Cdk7 pathway restoration in tumor cells with silenced or reduced CDK7 expression.

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