GRIP-1 CRISPR Activation Plasmid (h): sc-401800-ACT

NCOA2 encodes GRIP-1 (glucocorticoid receptor-interacting protein 1), a nuclear receptor coactivator that scaffolds transcriptional complexes and coordinates chromatin remodeling to regulate hormone-responsive gene expression. GRIP-1 integrates signaling from steroid and other nuclear receptors with transcriptional programs controlling proliferation, differentiation, metabolism, and inflammatory responses. It functions within coactivator assemblies that couple receptor binding to recruitment of histone acetyltransferases and mediator components, shaping promoter/enhancer activity. Dysregulation of NCOA2-dependent transcription has been linked to altered endocrine signaling and is implicated in oncogenic transcriptional rewiring and developmental pathology contexts.

GRIP-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NCOA2 expression without altering the underlying DNA sequence.

GRIP-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NCOA2 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 NCOA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GRIP-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NCOA2 locus and enabling the study of GRIP-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GRIP-1 pathway restoration in tumor cells with silenced or reduced NCOA2 expression.

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