PMS1 CRISPR Activation Plasmid (h): sc-418473-ACT

Human PMS1 encodes a MutL homolog that participates in DNA mismatch repair, supporting replication fidelity by coordinating recognition and processing of base–base mismatches and insertion–deletion loops. PMS1 functions within the MutL complex network to couple mismatch recognition to downstream excision and resynthesis, contributing to genome stability and suppression of spontaneous mutagenesis. Defects or altered regulation of mismatch repair genes can elevate microsatellite instability and increase mutation accumulation, processes broadly relevant to carcinogenesis and cellular stress responses. PMS1 is therefore studied in pathways governing DNA damage signaling, replication-associated repair, and maintenance of chromosomal integrity.

PMS1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PMS1 expression without altering the underlying DNA sequence.

PMS1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PMS1 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 PMS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PMS1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PMS1 locus and enabling the study of PMS1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PMS1 pathway restoration in tumor cells with silenced or reduced PMS1 expression.

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