CD73 Lentiviral Activation Particles (h): sc-400307-LAC

Human NT5E encodes CD73, a glycosylphosphatidylinositol-anchored ecto-5′-nucleotidase that dephosphorylates extracellular AMP to adenosine, thereby shaping purinergic signaling and nucleotide homeostasis at the cell surface. CD73-generated adenosine engages adenosine receptor pathways to regulate cAMP-dependent signaling, immune cell activation, endothelial barrier function, and vascular tone, and it collaborates functionally with upstream ectonucleotidases such as CD39 in ATP/ADP catabolism. NT5E/CD73 activity is frequently examined in the context of hypoxia- and inflammation-associated remodeling of the extracellular nucleotide milieu, where shifts in adenosine availability can alter leukocyte trafficking and stromal interactions. Dysregulated NT5E expression has been linked to immunoregulatory phenotypes in cancer microenvironments and to vascular and inflammatory pathobiology, supporting its use as a mechanistic node in studies of tissue stress responses.

CD73 Lentiviral Activation Particles (h) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient NT5E upregulation across a broader range of human cell types.

CD73 Lentiviral Activation Particles (h) 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 NT5E transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous CD73 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native NT5E 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.