Slfn14 CRISPR Activation Plasmid (h): sc-415573-ACT

Human SLFN14 encodes Schlafen family member 14 (Slfn14), a putative RNA-binding protein implicated in post-transcriptional regulation and control of protein synthesis in hematopoietic lineages. SLFN proteins are broadly linked to innate immune signaling and cellular stress responses, with roles that can influence ribosome-associated RNA handling and translational output. SLFN14 is particularly relevant to platelet biology, and altered SLFN14 function has been associated with inherited thrombocytopenia and platelet dysfunction phenotypes. These features make SLFN14 a useful node for studying gene expression control in megakaryocyte maturation, platelet production, and immune-adjacent regulatory programs.

Slfn14 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SLFN14 expression without altering the underlying DNA sequence.

Slfn14 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SLFN14 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 SLFN14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Slfn14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SLFN14 locus and enabling the study of Slfn14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Slfn14 pathway restoration in tumor cells with silenced or reduced SLFN14 expression.

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