5α-Reductase 1 CRISPR Activation Plasmid (h): sc-402151-ACT

SRD5A1 encodes human 5α-reductase 1, an NADPH-dependent microsomal oxidoreductase that catalyzes conversion of testosterone to the more potent androgen dihydrotestosterone (DHT). This enzymatic step is central to steroid hormone metabolism and androgen receptor signaling, influencing transcriptional programs that regulate proliferation, differentiation, and lipid homeostasis in androgen-responsive tissues. SRD5A1 activity is implicated in endocrine regulation and has been associated with androgen-driven disease biology, including altered steroidogenic flux in hormone-responsive cancers and dermatologic conditions. As a membrane-associated enzyme of the endoplasmic reticulum, 5α-reductase 1 also provides a tractable node for interrogating crosstalk between steroid biosynthesis, nuclear receptor signaling, and cellular metabolism.

5α-Reductase 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SRD5A1 expression without altering the underlying DNA sequence.

5α-Reductase 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SRD5A1 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 SRD5A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous 5α-Reductase 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SRD5A1 locus and enabling the study of 5α-Reductase 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of 5α-Reductase 1 pathway restoration in tumor cells with silenced or reduced SRD5A1 expression.

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