CYP21A2 Lentiviral Activation Particles (h): sc-402919-LAC

CYP21A2 encodes cytochrome P450 21-hydroxylase, an endoplasmic reticulum–associated monooxygenase that catalyzes key 21-hydroxylation steps in adrenal steroidogenesis, including conversion of progesterone and 17α-hydroxyprogesterone toward mineralocorticoid and glucocorticoid biosynthesis. This enzyme functions within heme-dependent P450 redox chemistry and is integrated with cholesterol-derived hormone pathways that shape adrenal zonation, endocrine feedback, and cellular metabolic state. Altered CYP21A2 activity is strongly linked to congenital adrenal hyperplasia phenotypes and provides a molecular entry point to study steroid precursor accumulation, disrupted cortisol/aldosterone production, and downstream signaling effects in relevant cell models. Because CYP21A2 resides in a complex genomic region near the highly homologous pseudogene CYP21A1P, precise interrogation of locus-specific regulation is often important for mechanistic studies.

CYP21A2 Lentiviral Activation Particles (h) address this need by packaging the complete synergistic activation mediator (SAM) transcriptional activation system into transduction-ready, high-titer lentiviral particles, enabling efficient CYP21A2 upregulation across a broader range of human cell types.

CYP21A2 Lentiviral Activation Particles (h) deliver all functional components of the synergistic activation mediator (SAM) system via lentiviral transduction. The system comprises three particle preparations co-transduced into target cells: one encoding catalytically inactive dCas9 (D10A and N863A mutations) fused to the VP64 transactivation domain with a blasticidin resistance gene; one encoding the MS2-p65-HSF1 fusion protein with a hygromycin resistance gene; and one encoding a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers with a puromycin resistance gene. Following lentiviral transduction and genomic integration of the expression cassettes, the SAM components are stably expressed and assemble at the target locus within the proximal promoter region upstream of the CYP21A2 transcriptional start site, where VP64, p65, and HSF1 act cooperatively to recruit endogenous transcriptional machinery and drive sustained upregulation of endogenous CYP21A2 expression. The use of nuclease-inactive dCas9 avoids the introduction of double-strand DNA breaks and preserves the native CYP21A2 genomic locus and regulatory architecture.

The lentiviral format offers several practical advantages: stable genomic integration supports heritable activation across cell divisions; high-titer particle preparations eliminate the need for in-house viral production; and compatibility with primary, non-dividing, and transfection-resistant cell types expands experimental accessibility. Successful transduction can be confirmed and enriched through triple antibiotic selection using puromycin, hygromycin, and blasticidin.

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