TASK-3 CRISPR Activation Plasmid (h): sc-403974-ACT

Human KCNK9 encodes TASK-3 (K2P9.1), a two-pore domain potassium channel that contributes to background “leak” K+ conductance and helps set resting membrane potential and membrane excitability. TASK-3 is sensitive to extracellular pH and modulates firing properties in excitable tissues, linking ion homeostasis to cellular signaling programs such as calcium-dependent pathways and activity-regulated transcription. Through its impact on membrane polarization, TASK-3 influences processes including neuronal and endocrine secretion, cell-cycle progression, and adaptation to metabolic or hypoxic stress. Altered KCNK9/TASK-3 expression or function has been associated with neurodevelopmental phenotypes and has been studied in the context of cancer biology, where potassium channel–dependent bioelectric signaling can affect proliferation and survival.

TASK-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KCNK9 expression without altering the underlying DNA sequence.

TASK-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KCNK9 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 KCNK9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TASK-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KCNK9 locus and enabling the study of TASK-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TASK-3 pathway restoration in tumor cells with silenced or reduced KCNK9 expression.

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