SF-1 CRISPR Activation Plasmid (h): sc-402334-ACT

NR5A1 encodes steroidogenic factor 1 (SF-1), an orphan nuclear receptor that functions as a master transcriptional regulator of adrenal and gonadal development, steroid hormone biosynthesis, and reproductive axis homeostasis. SF-1 coordinates gene networks controlling cholesterol transport and steroidogenic enzyme expression, integrating signaling inputs that shape endocrine differentiation programs. In human biology, altered NR5A1 activity is linked to disorders of sex development, impaired gonadal function, and adrenal insufficiency phenotypes, and it is also relevant to endocrine tumor biology. Because SF-1 modulates lineage specification and hormone-related transcriptional circuits, it is widely used to interrogate endocrine cell fate decisions, steroidogenesis pathways, and nuclear receptor cofactor dependencies.

SF-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NR5A1 expression without altering the underlying DNA sequence.

SF-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NR5A1 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 NR5A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NR5A1 locus and enabling the study of SF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SF-1 pathway restoration in tumor cells with silenced or reduced NR5A1 expression.

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