EGR1 CRISPR Activation Plasmid (m): sc-420133-ACT

Early growth response 1 (Egr1) encodes the zinc-finger transcription factor EGR1, an immediate-early gene rapidly induced by growth factors, stress signals, and neuronal activity. EGR1 regulates programs controlling cell cycle progression, apoptosis, differentiation, and synaptic plasticity by binding GC-rich promoter elements and coordinating chromatin and transcriptional responses. In mouse, EGR1 is a key node downstream of MAPK/ERK signaling and integrates inputs from calcium-dependent pathways and inflammatory cues to shape stimulus-dependent gene expression. Dysregulated EGR1 activity has been implicated in contexts including neuroadaptation, immune activation, fibrosis-related remodeling, and oncogenic transcriptional networks, making it useful for mechanistic studies of signal-to-transcription coupling.

EGR1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Egr1 expression without altering the underlying DNA sequence.

EGR1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Egr1 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 Egr1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EGR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Egr1 locus and enabling the study of EGR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EGR1 pathway restoration in tumor cells with silenced or reduced Egr1 expression.

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