Raf-1 Lentiviral Activation Particles (h): sc-400202-LAC

Human RAF1 encodes Raf-1, a serine/threonine kinase that functions as a central MAPK pathway node linking activated RAS family GTPases to MEK1/2 and ERK1/2 signaling. Raf-1 integrates cues from receptor tyrosine kinases and diverse scaffold proteins to regulate proliferation, differentiation, survival, and stress responses, with additional roles in cytoskeletal dynamics and mitochondrial signaling. Dysregulated RAF1 activity contributes to aberrant ERK output observed across multiple cancer-related signaling contexts and is also implicated in RASopathy phenotypes through pathway hyperactivation and altered developmental signaling. As such, RAF1 serves as a mechanistic entry point for studying pathway feedback, signal amplitude/duration control, and cross-talk with PI3K/AKT and apoptotic regulators.

Raf-1 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 RAF1 upregulation across a broader range of human cell types.

Raf-1 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 RAF1 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous Raf-1 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native RAF1 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.