RAP1 CRISPR Activation Plasmid (h): sc-401709-ACT

Human TERF2IP encodes RAP1, a shelterin-associated factor that binds TRF2 to regulate telomere protection, telomere length homeostasis, and suppression of aberrant DNA damage signaling at chromosome ends. Beyond telomeres, RAP1 influences transcriptional programs and chromatin organization, and has been linked to NF-κB-associated inflammatory signaling and broader genome stability pathways. Disruption of RAP1 function can alter telomere integrity, checkpoint responses, and cellular senescence-like phenotypes, making TERF2IP a relevant target for studies of genome maintenance mechanisms. Dysregulated telomere biology and RAP1-associated signaling networks are frequently investigated in the context of oncogenic transformation and age-associated cellular dysfunction without implying clinical outcomes.

RAP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TERF2IP expression without altering the underlying DNA sequence.

RAP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TERF2IP 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 TERF2IP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RAP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TERF2IP locus and enabling the study of RAP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RAP1 pathway restoration in tumor cells with silenced or reduced TERF2IP expression.

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