ADAR1 CRISPR Activation Plasmid (h): sc-401611-ACT

ADAR (ADAR1) encodes an RNA-editing enzyme that catalyzes adenosine-to-inosine (A-to-I) deamination within double-stranded RNA, reshaping transcript sequences, splice sites, and RNA structure. Through editing of endogenous dsRNA, ADAR1 modulates innate immune sensing pathways, including suppression of aberrant interferon signaling and regulation of cytosolic RNA surveillance. ADAR1 activity influences post-transcriptional gene regulation programs that impact cell-state transitions, stress responses, and tumor–immune interactions. Dysregulated RNA editing and perturbed ADAR1 function have been associated with interferonopathies and cancer-relevant phenotypes, making it a key target for mechanistic studies of RNA processing and immune regulation.

ADAR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAR expression without altering the underlying DNA sequence.

ADAR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAR 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 ADAR transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAR locus and enabling the study of ADAR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAR1 pathway restoration in tumor cells with silenced or reduced ADAR expression.

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