SAMD9 CRISPR Activation Plasmid (h): sc-418037-ACT

SAMD9 (sterile alpha motif domain containing 9) encodes a cytoplasmic protein implicated in innate immune regulation and control of cellular proliferation, with reported roles in restricting viral replication and modulating interferon-stimulated responses. SAMD9 function intersects with stress-response programs and homeostatic pathways that influence cell growth and survival, consistent with its proposed activity as a negative regulator of proliferation. Genetic alterations in SAMD9 are associated with inherited marrow failure and immunodeficiency phenotypes, and are recurrently implicated in pediatric myelodysplastic syndrome through germline or acquired variants. These links make SAMD9 a useful locus for studying inflammation-coupled growth control and mechanisms of somatic adaptation in hematopoietic cells.

SAMD9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SAMD9 expression without altering the underlying DNA sequence.

SAMD9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SAMD9 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 SAMD9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SAMD9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SAMD9 locus and enabling the study of SAMD9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SAMD9 pathway restoration in tumor cells with silenced or reduced SAMD9 expression.

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