Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
ATGL CRISPR Activation Plasmid (h): sc-401711-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ATGL CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- ATGL 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 ATGL CRISPR Activation Plasmid (h) and ATGL CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the PNPLA2 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: ATGL Antibody (F-7): sc-365278
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ATGL CRISPR Activation Plasmid (h) | sc-401711-ACT | 20 µg | $397.00 | |||
ATGL CRISPR Activation Plasmid (h2) | sc-401711-ACT-2 | 20 µg | $397.00 |
PNPLA2 encodes adipose triglyceride lipase (ATGL), the rate-limiting enzyme for the initial hydrolysis of triacylglycerol stored in cytosolic lipid droplets, generating diacylglycerol and free fatty acids for mitochondrial β-oxidation and cellular energy balance. ATGL activity integrates lipid droplet dynamics with nutrient-sensing and metabolic signaling networks, including PPAR-driven transcriptional programs and lipolytic regulation by cofactors such as ABHD5/CGI-58 and inhibitory proteins like G0S2. Dysregulated PNPLA2 expression or ATGL function perturbs triglyceride turnover and fatty acid flux, contributing to altered lipid storage, lipotoxic stress, and metabolic phenotypes observed across adipose, liver, heart, and skeletal muscle. As a central node in neutral lipid catabolism, PNPLA2/ATGL is widely studied in models of lipid storage disorders, insulin resistance-related pathways, and cellular responses to nutrient deprivation.
ATGL CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PNPLA2 expression without altering the underlying DNA sequence.
ATGL CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PNPLA2 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 PNPLA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATGL expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PNPLA2 locus and enabling the study of ATGL-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATGL pathway restoration in tumor cells with silenced or reduced PNPLA2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.