Ini1 CRISPR Activation Plasmid (m): sc-423027-ACT

Mouse Smarcb1 encodes Ini1 (also known as SNF5/BAF47), a core subunit of the SWI/SNF (BAF) ATP-dependent chromatin remodeling complex that regulates nucleosome positioning and transcriptional programs controlling cell cycle progression, lineage specification, and differentiation. Ini1 helps coordinate enhancer and promoter accessibility, integrating signaling inputs with chromatin state to modulate gene expression networks involved in development and DNA damage responses. Disruption of SMARCB1/INI1 function is linked to altered epigenetic regulation and aberrant proliferation, and loss of SWI/SNF subunits is a recurrent feature across multiple tumor types. As a central chromatin regulator, Smarcb1 is frequently studied in pathways governing transcriptional repression/activation balance, checkpoint control, and chromatin architecture.

Ini1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Smarcb1 expression without altering the underlying DNA sequence.

Ini1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Smarcb1 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 Smarcb1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Ini1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Smarcb1 locus and enabling the study of Ini1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Ini1 pathway restoration in tumor cells with silenced or reduced Smarcb1 expression.

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