RecQL5 CRISPR Activation Plasmid (h): sc-406407-ACT

Human RECQL5 encodes the RecQ helicase RecQL5, a genome maintenance factor that regulates DNA replication and transcription to limit aberrant recombination. RecQL5 interacts with RNA polymerase II and participates in DNA damage response pathways that promote replication fork stability, resolution of DNA secondary structures, and suppression of homologous recombination–associated genome rearrangements. By preserving chromosome integrity, RECQL5 supports accurate cell-cycle progression and counters accumulation of mutations linked to genomic instability phenotypes. Dysregulation of RECQL5 activity has been connected to elevated DNA damage signaling and altered recombination control, making it relevant for studies of genome instability mechanisms in disease models.

RecQL5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RECQL5 expression without altering the underlying DNA sequence.

RecQL5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RECQL5 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 RECQL5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RecQL5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RECQL5 locus and enabling the study of RecQL5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RecQL5 pathway restoration in tumor cells with silenced or reduced RECQL5 expression.

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