IRE1α Lentiviral Activation Particles (h): sc-400576-LAC

ERN1 encodes inositol-requiring enzyme 1 alpha (IRE1α), an ER-resident transmembrane kinase/endoribonuclease that functions as a primary sensor of unfolded proteins and a key initiator of the unfolded protein response (UPR). Upon ER stress, IRE1α oligomerizes and autophosphorylates, activating RNase outputs that include unconventional splicing of XBP1 mRNA to produce the transcription factor XBP1s and regulated IRE1-dependent decay (RIDD) of select transcripts to remodel secretory capacity. These activities integrate with proteostasis, lipid metabolism, autophagy, and inflammatory signaling via UPR crosstalk with pathways such as NF-κB and JNK. Dysregulated ERN1/IRE1α signaling is linked to cellular stress adaptation in cancer, metabolic disease contexts, neurodegeneration, and immune cell differentiation, making it a widely used node for mechanistic studies of ER stress signaling.

IRE1α 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 ERN1 upregulation across a broader range of human cell types.

IRE1α 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 ERN1 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous IRE1α expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native ERN1 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.