SMC3 CRISPR Activation Plasmid (h): sc-403950-ACT

SMC3 encodes a core structural maintenance of chromosomes protein that forms the cohesin complex with SMC1A and associated subunits to establish sister chromatid cohesion during S phase and ensure accurate chromosome segregation. Beyond mitosis, SMC3-dependent cohesin dynamics regulate DNA double-strand break repair, replication fork stability, and higher-order genome organization that influences enhancer–promoter communication and transcriptional programs. These functions place SMC3 at the center of pathways controlling genome stability and cell-cycle progression, with dysregulation linked to chromosomal instability phenotypes and cohesin-related developmental disorders. In cancer biology, altered cohesin activity involving SMC3 is frequently studied for its effects on aneuploidy, DNA repair capacity, and transcriptional rewiring.

SMC3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMC3 expression without altering the underlying DNA sequence.

SMC3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMC3 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 SMC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMC3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMC3 locus and enabling the study of SMC3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMC3 pathway restoration in tumor cells with silenced or reduced SMC3 expression.

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