p14ARF/p16 Lentiviral Activation Particles (h): sc-400018-LAC

CDKN2A encodes two tumor suppressor proteins, p16INK4a and p14ARF, produced from alternative reading frames that constrain cell-cycle progression and coordinate senescence programs. p16INK4a inhibits CDK4/6 to maintain RB1 in a hypophosphorylated state, limiting E2F-dependent S-phase entry, while p14ARF stabilizes TP53 by antagonizing MDM2 and promoting cell-cycle arrest or apoptosis in response to oncogenic stress. Through these RB and p53 axis connections, CDKN2A integrates mitogenic signaling with checkpoint control, DNA damage responses, and aging-associated transcriptional changes. Altered CDKN2A regulation is frequently linked to dysregulated proliferation, impaired senescence, and tumorigenic phenotypes, making it a central node for mechanistic studies in cancer biology and cell fate control.

p14ARF/p16 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 CDKN2A upregulation across a broader range of human cell types.

p14ARF/p16 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 CDKN2A transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous p14ARF/p16 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native CDKN2A 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.