SMC1 CRISPR Activation Plasmid (h): sc-403715-ACT

SMC1B encodes a meiosis-associated structural maintenance of chromosomes (SMC) subunit that partners with cohesin components to regulate sister chromatid cohesion, chromosome synapsis, and accurate segregation during gametogenesis. Cohesin-mediated higher-order chromatin organization influences DNA replication timing and DNA damage responses, linking SMC family activity to genome stability pathways and transcriptional regulation. Dysregulation of cohesin biology is broadly implicated in aneuploidy, infertility-related mechanisms, and cohesinopathies, and altered cohesin function is also relevant to cancer-associated chromosomal instability. In human cell models, modulating SMC1B expression supports studies of chromatin architecture, recombination-related processes, and cell cycle control.

SMC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMC1B expression without altering the underlying DNA sequence.

SMC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMC1B 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 SMC1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMC1B locus and enabling the study of SMC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMC1 pathway restoration in tumor cells with silenced or reduced SMC1B expression.

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