SMC4 CRISPR Activation Plasmid (h): sc-405067-ACT

SMC4 encodes a core structural maintenance of chromosomes ATPase that, together with SMC2, forms the catalytic scaffold of the condensin complexes required for mitotic chromosome condensation, sister chromatid resolution, and faithful chromosome segregation. Through ATP-dependent DNA loop extrusion and higher-order chromatin organization, SMC4 supports replication-associated chromatin dynamics, DNA damage response coordination, and maintenance of genome stability. Dysregulated SMC4 expression or function has been associated with chromosomal instability and aberrant proliferation phenotypes observed across multiple tumor contexts, making it a useful node for studying aneuploidy and cell-cycle checkpoint control. In addition, SMC4-dependent chromatin architecture influences transcriptional programs linked to mitosis and stress responses, providing a mechanistic bridge between chromosome structure and gene regulation.

SMC4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SMC4 expression without altering the underlying DNA sequence.

SMC4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SMC4 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 SMC4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SMC4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SMC4 locus and enabling the study of SMC4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SMC4 pathway restoration in tumor cells with silenced or reduced SMC4 expression.

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