OAS1 CRISPR Activation Plasmid (h): sc-402414-ACT

Human OAS1 (2′-5′-oligoadenylate synthetase 1) is an interferon-stimulated enzyme that detects viral double-stranded RNA and catalyzes synthesis of 2′-5′-linked oligoadenylates. These second messengers activate RNase L, promoting degradation of viral and cellular RNA and shaping innate immune signaling, including feedback into type I interferon pathways. OAS1 activity influences antiviral restriction, inflammatory tone, and stress responses, and genetic or expression variation has been associated with differential susceptibility and outcomes in viral infection contexts and interferon-driven immune phenotypes. As a central node in RNA-sensing immunity, OAS1 is widely studied for its impact on host–pathogen interactions and cytokine-regulated transcriptional programs.

OAS1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous OAS1 expression without altering the underlying DNA sequence.

OAS1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the OAS1 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 OAS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OAS1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native OAS1 locus and enabling the study of OAS1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OAS1 pathway restoration in tumor cells with silenced or reduced OAS1 expression.

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