E2F1 CRISPR Activation Plasmid (h): sc-400123-ACT

E2F1 (E2F transcription factor 1) is a sequence-specific transcription factor that governs G1/S transition by coordinating expression of genes required for DNA replication, nucleotide metabolism, and cell-cycle progression. Its activity is tightly regulated by the RB/E2F axis, integrating mitogenic signaling with checkpoint control, and it intersects with p53-dependent apoptosis and DNA damage responses. Dysregulated E2F1 signaling is frequently linked to uncontrolled proliferation, replication stress, and genomic instability observed across cancer and other proliferative disorders. Because E2F1 can also influence apoptosis, senescence, and differentiation programs, it is widely used to probe context-dependent cell fate decisions.

E2F1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous E2F1 expression without altering the underlying DNA sequence.

E2F1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the E2F1 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 E2F1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous E2F1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native E2F1 locus and enabling the study of E2F1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of E2F1 pathway restoration in tumor cells with silenced or reduced E2F1 expression.

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