TRF2 CRISPR Activation Plasmid (h): sc-401289-ACT

Human TERF2 encodes telomeric repeat-binding factor 2 (TRF2), a core shelterin component that binds double-stranded telomeric DNA to promote t-loop formation and protect chromosome ends from being recognized as DNA double-strand breaks. TRF2 suppresses ATM-dependent DNA damage signaling and limits end-to-end chromosomal fusions by preventing inappropriate non-homologous end joining at telomeres, thereby supporting genome stability and proper cell-cycle progression. Through its roles in telomere capping, replication, and telomere length homeostasis, TERF2 helps coordinate telomere biology with DNA repair and checkpoint pathways. Dysregulation of TRF2 has been linked to telomere dysfunction phenotypes and is frequently studied in contexts of genomic instability associated with aging-related processes and cancer biology.

TRF2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TERF2 expression without altering the underlying DNA sequence.

TRF2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TERF2 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 TERF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRF2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TERF2 locus and enabling the study of TRF2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRF2 pathway restoration in tumor cells with silenced or reduced TERF2 expression.

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