NEDD4 Lentiviral Activation Particles (m): sc-421848-LAC

Mouse Nedd4 encodes NEDD4, a HECT-domain E3 ubiquitin ligase that regulates protein turnover and signaling by catalyzing ubiquitin transfer to membrane receptors, ion channels, and signaling intermediates. Through WW-domain recognition of PY-motif substrates, NEDD4 influences endocytosis and trafficking of surface proteins and modulates pathways such as PI3K–AKT, TGF-β, and EGFR-associated signaling, with downstream effects on cell growth, survival, and migration. NEDD4 activity also intersects with proteostasis and stress-response networks, shaping ubiquitin-dependent remodeling of cellular signaling complexes. Dysregulated NEDD4-dependent ubiquitination has been implicated in disease-relevant phenotypes including altered epithelial transport, aberrant proliferative signaling, and immune and neuronal dysfunction, making it a useful node for mechanistic studies.

NEDD4 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 Nedd4 upregulation across a broader range of human cell types.

NEDD4 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 Nedd4 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous NEDD4 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native Nedd4 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.