RNF8 Lentiviral Activation Particles (h): sc-401909-LAC

Human RNF8 encodes a RING finger E3 ubiquitin ligase that orchestrates the DNA damage response by catalyzing ubiquitin signaling at sites of DNA double-strand breaks. Through crosstalk with ATM/ATR pathways, RNF8 promotes recruitment and retention of key repair factors including 53BP1, BRCA1, and downstream ubiquitin-dependent assemblies that regulate checkpoint activation and repair pathway choice. RNF8 is also implicated in chromatin remodeling and maintenance of genomic stability, linking its activity to cellular outcomes such as cell-cycle arrest, apoptosis, and senescence under genotoxic stress. Dysregulated RNF8-dependent ubiquitination has been associated with genome instability phenotypes and altered damage signaling observed across multiple cancer-relevant contexts and other disorders of DNA repair.

RNF8 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 RNF8 upregulation across a broader range of human cell types.

RNF8 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 RNF8 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous RNF8 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native RNF8 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.