LGP2 CRISPR Activation Plasmid (h): sc-406991-ACT

DHX58 encodes LGP2, a DExD/H-box RNA helicase–like sensor that modulates cytosolic detection of viral RNA and shapes innate antiviral signaling. LGP2 interacts with RIG-I–like receptor pathways, influencing MAVS-dependent induction of type I interferons and interferon-stimulated genes, and can tune the magnitude and duration of inflammatory transcriptional programs. By regulating RNA sensing and downstream NF-κB and IRF-driven responses, DHX58 contributes to cellular immune homeostasis and pathogen restriction. Dysregulated DHX58/LGP2 activity and altered interferon signaling have been associated with susceptibility to viral infection, chronic inflammation, and interferon-related immune phenotypes relevant to immunology and host–pathogen research.

LGP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DHX58 expression without altering the underlying DNA sequence.

LGP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DHX58 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 DHX58 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LGP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DHX58 locus and enabling the study of LGP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LGP2 pathway restoration in tumor cells with silenced or reduced DHX58 expression.

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