PYGL CRISPR Activation Plasmid (h): sc-401950-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PYGL CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- PYGL 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 PYGL CRISPR Activation Plasmid (h) and PYGL CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the PYGL transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PYGL CRISPR Activation Plasmid (h) | sc-401950-ACT | 20 µg | $397.00 |
Human PYGL encodes the liver isoform of glycogen phosphorylase, a key cytosolic enzyme that catalyzes phosphorolytic cleavage of glycogen to generate glucose-1-phosphate for downstream glycolysis and maintenance of systemic glucose homeostasis. PYGL activity integrates hormonal and nutrient signaling with glycogenolysis and broader carbohydrate metabolism, contributing to cellular energy balance under fasting and metabolic stress. Dysregulation of hepatic glycogen breakdown is linked to inborn errors of glycogen metabolism and can influence metabolic phenotypes associated with insulin resistance and liver function. As a pathway node connecting glycogen storage to central carbon metabolism, PYGL is widely studied in hepatic biology, metabolic flux regulation, and energy-sensing networks.
PYGL CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PYGL expression without altering the underlying DNA sequence.
PYGL CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PYGL 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 PYGL transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PYGL expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PYGL locus and enabling the study of PYGL-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PYGL pathway restoration in tumor cells with silenced or reduced PYGL expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.