FANCD2 Lentiviral Activation Particles (h): sc-400445-LAC

FANCD2 encodes a central effector of the Fanconi anemia (FA) DNA interstrand crosslink repair pathway and coordinates genome maintenance during S phase. Upon replication stress or crosslink damage, FANCD2 is monoubiquitinated and accumulates at stalled forks, where it scaffolds repair with partners including FANCI, BRCA1/2, RAD51, and nucleases that process DNA lesions. This activity interfaces with homologous recombination, checkpoint signaling, and replication fork protection, linking FANCD2 function to chromosomal stability and cell-cycle progression. Defects in FANCD2-mediated repair are associated with Fanconi anemia phenotypes and increased sensitivity to crosslinking agents, making the pathway a key focus in studies of DNA damage responses and cancer-associated genome instability.

FANCD2 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 FANCD2 upregulation across a broader range of human cell types.

FANCD2 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 FANCD2 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous FANCD2 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native FANCD2 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.