ADAR2 CRISPR Activation Plasmid (h): sc-401166-ACT

Human ADARB1 encodes ADAR2, an adenosine deaminase acting on RNA that catalyzes site-selective A-to-I RNA editing within double-stranded RNA structures. This post-transcriptional modification can recode codons, alter splice sites, and modulate RNA stability, thereby shaping neuronal excitability and synaptic signaling through editing of neurotransmission-associated transcripts. ADAR2 activity intersects with innate immune RNA-sensing pathways by reducing the immunogenicity of endogenous dsRNA and influencing interferon-stimulated gene programs. Dysregulated RNA editing patterns involving ADAR2 have been linked to neurological and neurodevelopmental phenotypes and are studied as molecular contributors to disease-associated transcriptome remodeling.

ADAR2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADARB1 expression without altering the underlying DNA sequence.

ADAR2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADARB1 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 ADARB1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAR2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADARB1 locus and enabling the study of ADAR2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAR2 pathway restoration in tumor cells with silenced or reduced ADARB1 expression.

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