ANO4 Lentiviral Activation Particles (h2): sc-406887-LAC-2

Human ANO4 encodes anoctamin-4, a member of the TMEM16/anoctamin family of multi-pass membrane proteins implicated in calcium-regulated ion transport and phospholipid scrambling at cellular membranes. ANO4 activity is linked to regulation of membrane potential, intracellular Ca²⁺-dependent signaling, and remodeling events that influence vesicular trafficking, apoptosis, and cellular excitability. Altered expression or dysfunction of TMEM16 family proteins, including ANO4, has been associated with neurological and other complex phenotypes, supporting its investigation in mechanisms of neurodegeneration and tissue homeostasis. ANO4 gene editing and functional perturbation studies enable pathway dissection in model systems, including electrophysiology and lipid-scrambling assays, to map downstream signaling networks and genotype–phenotype relationships.

ANO4 Lentiviral Activation Particles (h2) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient ANO4 upregulation across a broader range of human cell types.

ANO4 Lentiviral Activation Particles (h2) 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 ANO4 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous ANO4 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native ANO4 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.