DP-1 CRISPR Activation Plasmid (h): sc-403331-ACT

Human TFDP1 encodes DP-1, a sequence-specific transcription factor that heterodimerizes with E2F family members to regulate genes required for G1/S transition, DNA replication, and mitotic progression. DP-1/E2F complexes integrate signals from the RB pathway to coordinate cell-cycle entry, checkpoint control, and proliferation-associated transcriptional programs. Through control of replication licensing and S-phase gene expression, TFDP1 activity influences genome stability and cellular responses to oncogenic stress. Dysregulated TFDP1–E2F signaling is frequently implicated in proliferation-driven disease biology, making DP-1 a useful node for studying cell-cycle control, DNA damage responses, and transcriptional network rewiring in human cells.

DP-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TFDP1 expression without altering the underlying DNA sequence.

DP-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TFDP1 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 TFDP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DP-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TFDP1 locus and enabling the study of DP-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DP-1 pathway restoration in tumor cells with silenced or reduced TFDP1 expression.

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