SMARCA4/Brg1 CRISPR Activation Plasmid (h): sc-400168-ACT

SMARCA4 (Brg1) encodes the ATPase core of the SWI/SNF (BAF) chromatin-remodeling complex, which repositions nucleosomes to regulate transcriptional programs controlling cell identity, proliferation, and differentiation. Through ATP-dependent remodeling, SMARCA4 influences enhancer accessibility and coordinates with lineage-defining transcription factors to modulate RNA polymerase II activity. It participates in processes such as DNA damage signaling, replication stress responses, and cell-cycle checkpoint control by shaping chromatin architecture at key regulatory loci. Dysregulation or loss of SMARCA4 function is recurrent in multiple tumor contexts, linking altered SWI/SNF activity to epigenetic reprogramming, genomic instability, and aberrant developmental pathway output.

SMARCA4/Brg1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMARCA4 expression without altering the underlying DNA sequence.

SMARCA4/Brg1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMARCA4 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 SMARCA4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMARCA4/Brg1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMARCA4 locus and enabling the study of SMARCA4/Brg1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMARCA4/Brg1 pathway restoration in tumor cells with silenced or reduced SMARCA4 expression.

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