Slfn5 CRISPR Activation Plasmid (h): sc-408333-ACT

Human SLFN5 (Schlafen family member 5) encodes Slfn5, a nuclear interferon-stimulated protein that modulates transcriptional programs linked to innate immune signaling and cell-state regulation. Slfn5 has been implicated in controlling interferon response gene networks, influencing chromatin-associated processes and transcription factor activity that shape antiviral and inflammatory outputs. Altered SLFN5 expression has been associated with tumor cell proliferation, migration, and immune microenvironment signaling in multiple cancer contexts, supporting its utility as a mechanistic node connecting interferon pathways to oncogenic phenotypes. SLFN5 is therefore widely studied in models of infection, inflammation, and cancer biology to dissect how interferon-driven transcriptional repression or activation impacts cellular fitness.

Slfn5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLFN5 expression without altering the underlying DNA sequence.

Slfn5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLFN5 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the SLFN5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Slfn5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLFN5 locus and enabling the study of Slfn5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Slfn5 pathway restoration in tumor cells with silenced or reduced SLFN5 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.