SRCAP CRISPR Activation Plasmid (h): sc-403049-ACT

SRCAP (Snf2-related CREBBP activator protein) encodes an ATP-dependent chromatin remodeler that serves as the catalytic core of the SRCAP complex, which catalyzes exchange of canonical H2A for the histone variant H2A.Z at promoters and enhancers. Through H2A.Z deposition, SRCAP helps coordinate transcriptional programs linked to cell cycle progression, DNA damage responses, and lineage-specific differentiation by modulating nucleosome positioning and chromatin accessibility. SRCAP also interfaces functionally with CBP/CREBBP-associated transcriptional regulation and broader epigenetic control of gene expression. Dysregulation or mutation of SRCAP has been associated with developmental disorders and altered transcriptional homeostasis, motivating mechanistic studies of chromatin remodeling in disease-relevant cell states.

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

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

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