BAF60a CRISPR Activation Plasmid (h): sc-402641-ACT

SMARCD1 encodes BAF60a, a core subunit of the SWI/SNF (BAF) ATP-dependent chromatin remodeling complex that regulates nucleosome positioning and chromatin accessibility to control transcriptional programs. BAF60a coordinates interactions between transcription factors and the remodeling machinery to influence lineage specification, cell cycle progression, and stimulus-responsive gene expression, including pathways linked to development and differentiation. As part of the BAF complex, SMARCD1 contributes to epigenetic regulation of enhancer and promoter states and intersects with signaling networks such as hormone receptor and stress-response transcriptional circuits. Dysregulation of SWI/SNF components is associated with altered chromatin states observed across multiple cancers and neurodevelopmental phenotypes, making BAF60a a relevant target for mechanistic studies of transcriptional control.

BAF60a CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMARCD1 expression without altering the underlying DNA sequence.

BAF60a CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMARCD1 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 SMARCD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BAF60a expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMARCD1 locus and enabling the study of BAF60a-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BAF60a pathway restoration in tumor cells with silenced or reduced SMARCD1 expression.

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