SRC-1 CRISPR Activation Plasmid (h): sc-400879-ACT

NCOA1 encodes steroid receptor coactivator-1 (SRC-1), a transcriptional coactivator that integrates ligand-dependent signaling from nuclear receptors and other transcription factors to modulate chromatin accessibility and RNA polymerase II–driven gene expression. SRC-1 recruits histone acetyltransferases and additional co-regulators to coordinate programs involved in hormone-responsive transcription, metabolic homeostasis, cell-cycle control, and developmental differentiation. Through crosstalk with estrogen receptor, androgen receptor, glucocorticoid receptor, and growth factor–responsive pathways, SRC-1 contributes to context-dependent transcriptional remodeling. Altered NCOA1/SRC-1 expression or coactivator activity has been linked to dysregulated endocrine signaling and transcriptional networks observed in hormone-associated disease biology, providing a mechanistic entry point for studying pathway-specific gene regulation.

SRC-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCOA1 expression without altering the underlying DNA sequence.

SRC-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCOA1 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 NCOA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SRC-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCOA1 locus and enabling the study of SRC-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SRC-1 pathway restoration in tumor cells with silenced or reduced NCOA1 expression.

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