ASIC3 CRISPR Activation Plasmid (h): sc-405106-ACT

Human ASIC3 (acid-sensing ion channel subunit 3) is a proton-gated, amiloride-sensitive cation channel of the DEG/ENaC family that opens in response to extracellular acidification, enabling Na⁺ influx and membrane depolarization. It is highly relevant to sensory neuron excitability and transduces changes in tissue pH associated with inflammation, ischemia, and mechanical stress, integrating with broader ion homeostasis and excitatory signaling processes. ASIC3 activity influences calcium-dependent downstream signaling indirectly through depolarization-driven pathways and contributes to nociceptive and mechanosensory circuits. Dysregulated ASIC3 expression or function has been linked in research settings to pain hypersensitivity, inflammatory responses, and neuromuscular or musculoskeletal physiology under acidic microenvironments.

ASIC3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ASIC3 expression without altering the underlying DNA sequence.

ASIC3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ASIC3 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 ASIC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ASIC3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ASIC3 locus and enabling the study of ASIC3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ASIC3 pathway restoration in tumor cells with silenced or reduced ASIC3 expression.

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