serum reponse factor /SRF CRISPR Activation Plasmid (h): sc-400470-ACT

Human SRF (serum reponse factor) is a MADS-box transcription factor that binds serum response elements to coordinate immediate-early gene expression programs in response to growth factors, mitogens, and mechanical cues. It integrates signaling through RhoA–actin dynamics via MRTF cofactors and MAPK/ERK pathways via ternary complex factors to regulate cytoskeletal organization, cell adhesion, migration, and myogenic differentiation. SRF-dependent transcriptional networks shape smooth and cardiac muscle gene expression and broader stress-responsive remodeling processes. Dysregulated SRF activity has been associated with aberrant proliferative signaling, altered motility, and pathological tissue remodeling, making it a useful node for mechanistic studies of gene regulatory circuitry.

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

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

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