HDGFRP3 CRISPR Activation Plasmid (h2): sc-416344-ACT-2

Human HDGFRP3 (hepatoma-derived growth factor-related protein 3) encodes a nuclear, chromatin-associated protein implicated in regulation of transcription and chromatin dynamics through interactions with DNA and nucleic acid–binding partners. HDGFRP3 is linked to processes such as cell cycle control, proliferation, and differentiation, and it is frequently studied in the context of epigenetic regulation and gene expression programs in transformed cells. Altered HDGFRP3 expression has been reported across multiple cancer types, supporting its use as a molecular readout in oncology-focused pathway studies without implying clinical utility. Gene editing of HDGFRP3 in human cell models enables mechanistic dissection of chromatin-dependent signaling networks, mapping of downstream transcriptional targets by RNA-seq/ChIP-based assays, and functional screens probing growth, stress responses, and genome regulation.

HDGFRP3 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous HDGFRP3 expression without altering the underlying DNA sequence.

HDGFRP3 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HDGFRP3 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 HDGFRP3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HDGFRP3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HDGFRP3 locus and enabling the study of HDGFRP3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HDGFRP3 pathway restoration in tumor cells with silenced or reduced HDGFRP3 expression.

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