ARID1B CRISPR Activation Plasmid (h): sc-402365-ACT

ARID1B encodes AT-rich interaction domain-containing protein 1B, an essential subunit of the SWI/SNF (BAF) ATP-dependent chromatin remodeling complex that regulates nucleosome positioning and transcriptional programs during development and differentiation. By coordinating enhancer accessibility and promoter remodeling, ARID1B influences lineage specification, cell-cycle control, and DNA damage-responsive gene expression across multiple signaling contexts. Altered ARID1B function has been implicated in neurodevelopmental disorders and in diverse cancers where disrupted chromatin regulation contributes to aberrant transcriptional states and impaired cellular identity. These properties make ARID1B a widely studied node in epigenetic regulation, transcriptional control, and genotype-to-phenotype mapping.

ARID1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARID1B expression without altering the underlying DNA sequence.

ARID1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARID1B 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 ARID1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARID1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARID1B locus and enabling the study of ARID1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARID1B pathway restoration in tumor cells with silenced or reduced ARID1B expression.

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