mSin3B CRISPR Activation Plasmid (h): sc-401638-ACT

Human SIN3B encodes mSin3B, a core component of the SIN3 transcriptional corepressor complex that scaffolds histone deacetylases (including HDAC1/2) and associated chromatin regulators to shape epigenetic states. Through interactions with sequence-specific transcription factors and chromatin-binding proteins, mSin3B modulates gene programs governing cell-cycle control, differentiation, senescence, and stress-responsive transcription. SIN3B-dependent chromatin remodeling influences transcriptional repression and lineage commitment pathways, linking its activity to mechanisms relevant to oncogenic transcriptional dysregulation and altered cellular proliferation states. Because Sin3/HDAC assemblies integrate multiple signaling inputs, SIN3B is frequently studied in contexts where epigenetic repression and transcriptional network stability are perturbed.

mSin3B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SIN3B expression without altering the underlying DNA sequence.

mSin3B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SIN3B 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 SIN3B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous mSin3B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SIN3B locus and enabling the study of mSin3B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of mSin3B pathway restoration in tumor cells with silenced or reduced SIN3B expression.

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