Rad21 CRISPR Activation Plasmid (m): sc-422577-ACT

Mouse RAD21 is a core component of the cohesin complex that stabilizes sister chromatid cohesion and coordinates chromosome segregation during mitosis and meiosis. Beyond its structural role, RAD21 contributes to DNA damage repair, replication fork stability, and 3D genome organization by shaping chromatin loop dynamics and long-range enhancer–promoter interactions. Through these functions, Rad21 helps regulate transcriptional programs controlling proliferation and differentiation, linking cohesin activity to developmental patterning and tissue homeostasis. Dysregulated cohesin function, including altered RAD21 levels, has been associated with genome instability and aberrant gene expression signatures relevant to cancer biology and cohesin-related developmental syndromes.

Rad21 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Rad21 expression without altering the underlying DNA sequence.

Rad21 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Rad21 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 Rad21 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rad21 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Rad21 locus and enabling the study of Rad21-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rad21 pathway restoration in tumor cells with silenced or reduced Rad21 expression.

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