SMC1α CRISPR Activation Plasmid (h): sc-402865-ACT

SMC1A encodes structural maintenance of chromosomes protein 1α (SMC1α), a core component of the cohesin complex that controls sister chromatid cohesion, chromosome segregation, and higher-order chromatin organization. Through cohesin-dependent DNA loop extrusion and coordination with CTCF, SMC1α influences transcriptional regulation, DNA replication timing, and maintenance of genome stability. SMC1α also participates in DNA damage signaling and repair, supporting checkpoint responses to replication stress and double-strand breaks. Dysregulation or mutation of SMC1A and cohesin pathway components is linked to cohesinopathies and has been implicated in genome instability and altered gene expression programs relevant to cancer biology.

SMC1α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMC1A expression without altering the underlying DNA sequence.

SMC1α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMC1A 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 SMC1A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMC1α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMC1A locus and enabling the study of SMC1α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMC1α pathway restoration in tumor cells with silenced or reduced SMC1A expression.

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