Von Hippel Lindau/VHL CRISPR Activation Plasmid (h): sc-400528-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Von Hippel Lindau/VHL CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Von Hippel Lindau/VHL CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by Von Hippel Lindau/VHL CRISPR Activation Plasmid (h) and Von Hippel Lindau/VHL CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the VHL transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Von Hippel Lindau/VHL Antibody (VHL40): sc-135657
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Von Hippel Lindau/VHL CRISPR Activation Plasmid (h) | sc-400528-ACT | 20 µg | $397.00 | |||
Von Hippel Lindau/VHL CRISPR Activation Plasmid (h2) | sc-400528-ACT-2 | 20 µg | $397.00 |
VHL encodes the von Hippel–Lindau tumor suppressor, the substrate recognition component of the VCB-CUL2 E3 ubiquitin ligase complex that targets hydroxylated HIF-1α and HIF-2α for proteasomal degradation. Through oxygen-dependent regulation of HIF transcriptional programs, VHL controls hypoxia signaling, angiogenic factor expression, metabolic adaptation, and erythropoiesis-related pathways. VHL also contributes to extracellular matrix assembly, microtubule stability, and cilia-associated signaling, linking it to broader regulation of cell polarity and stress responses. Loss or dysfunction of VHL is strongly associated with aberrant hypoxic signaling and is a hallmark of Von Hippel–Lindau disease and renal cell carcinoma biology.
Von Hippel Lindau/VHL CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VHL expression without altering the underlying DNA sequence.
Von Hippel Lindau/VHL CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VHL 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 VHL transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Von Hippel Lindau/VHL expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VHL locus and enabling the study of Von Hippel Lindau/VHL-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Von Hippel Lindau/VHL pathway restoration in tumor cells with silenced or reduced VHL expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.