RCC1 CRISPR Activation Plasmid (h): sc-402078-ACT

RCC1 (regulator of chromosome condensation 1) encodes a chromatin-bound guanine nucleotide exchange factor that generates Ran-GTP on chromosomes, establishing the Ran gradient required for mitotic spindle assembly and nuclear envelope reformation. By controlling Ran-dependent nucleocytoplasmic transport, RCC1 coordinates cell cycle progression, chromosome condensation, and post-mitotic nuclear organization. RCC1 function intersects with DNA replication and genome stability programs through its coupling to chromatin dynamics and mitotic control. Dysregulated RCC1 activity or expression has been associated with proliferative phenotypes and altered checkpoint responses, making it relevant for studying mechanisms that support oncogenic growth and aneuploidy.

RCC1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RCC1 expression without altering the underlying DNA sequence.

RCC1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RCC1 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 RCC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RCC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RCC1 locus and enabling the study of RCC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RCC1 pathway restoration in tumor cells with silenced or reduced RCC1 expression.

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