NKCC1 Lentiviral Activation Particles (m): sc-422971-LAC

Slc12a2 encodes the Na⁺-K⁺-2Cl⁻ cotransporter NKCC1, a widely expressed electroneutral transporter that couples sodium and potassium gradients to chloride influx, thereby regulating intracellular Cl⁻ homeostasis, cell volume, and epithelial fluid secretion. In mouse tissues, NKCC1 contributes to ion and water movement across epithelia, influences neuronal excitability through chloride-dependent GABAergic signaling, and supports secretory and osmotic stress responses. NKCC1 activity integrates with WNK–SPAK/OSR1 kinase signaling and interacts functionally with KCC cotransporters to set cellular chloride levels. Dysregulated NKCC1-linked transport processes have been associated with edema and barrier dysfunction, altered neuronal network activity, and pathophysiology involving disrupted salt and water balance, making Slc12a2 a relevant target for mechanistic studies of transport biology.

NKCC1 Lentiviral Activation Particles (m) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient Slc12a2 upregulation across a broader range of human cell types.

NKCC1 Lentiviral Activation Particles (m) deliver all functional components of the synergistic activation mediator (SAM) system via lentiviral transduction. The system comprises three particle preparations co-transduced into target cells: one encoding catalytically inactive dCas9 (D10A and N863A mutations) fused to the VP64 transactivation domain with a blasticidin resistance gene; one encoding the MS2-p65-HSF1 fusion protein with a hygromycin resistance gene; and one encoding a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers with a puromycin resistance gene. Following lentiviral transduction and genomic integration of the expression cassettes, the SAM components are stably expressed and assemble at the target locus within the proximal promoter region upstream of the Slc12a2 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous NKCC1 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native Slc12a2 genomic locus and regulatory architecture.

The lentiviral format offers several practical advantages: stable genomic integration supports heritable activation across cell divisions; high-titer particle preparations eliminate the need for in-house viral production; and compatibility with primary, non-dividing, and transfection-resistant cell types expands experimental accessibility. Successful transduction can be confirmed and enriched through triple antibiotic selection using puromycin, hygromycin, and blasticidin.

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