Keap1 Lentiviral Activation Particles (h): sc-400190-LAC

Human KEAP1 encodes Keap1, a substrate adaptor for the CUL3–RBX1 E3 ubiquitin ligase complex that controls proteasomal turnover of NRF2 (NFE2L2) and thereby calibrates antioxidant and xenobiotic stress responses. By sensing electrophiles and reactive oxygen species through cysteine-rich domains, Keap1 regulates NRF2 nuclear accumulation and transcription of cytoprotective genes involved in glutathione metabolism, NADPH regeneration, and phase II detoxification. This KEAP1–NRF2 axis integrates redox homeostasis with metabolic rewiring, inflammatory signaling, and autophagy via interactions with proteins such as p62/SQSTM1. Dysregulation of KEAP1-dependent control of NRF2 signaling is frequently studied in contexts of oxidative stress biology, including carcinogenesis, chemoresistance-associated phenotypes, and chronic tissue injury models.

Keap1 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 KEAP1 upregulation across a broader range of human cell types.

Keap1 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 KEAP1 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous Keap1 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native KEAP1 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.