Elk-1 CRISPR Activation Plasmid (h): sc-400385-ACT

ELK1 encodes Elk-1, an ETS-domain transcription factor that functions as a nuclear effector of mitogen-activated signaling, most prominently downstream of ERK/MAPK. Upon phosphorylation, Elk-1 cooperates with serum response factor at serum response elements to regulate immediate early gene transcription programs that control proliferation, differentiation, synaptic plasticity, and cellular stress responses. ELK1 activity integrates cues from growth factor receptors and other kinase pathways to shape context-specific transcriptional outputs, making it a useful node for studying signal-dependent gene regulation. Dysregulated Elk-1 signaling and target gene expression have been implicated in oncogenic transcriptional programs and neurobiology-related phenotypes, supporting its relevance in mechanistic disease modeling.

Elk-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ELK1 expression without altering the underlying DNA sequence.

Elk-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ELK1 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 ELK1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Elk-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ELK1 locus and enabling the study of Elk-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Elk-1 pathway restoration in tumor cells with silenced or reduced ELK1 expression.

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