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.
TRC8 CRISPR Activation Plasmid (h): sc-403689-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TRC8 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- TRC8 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 TRC8 CRISPR Activation Plasmid (h) and TRC8 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the RNF139 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: TRC8 Antibody (H-9): sc-390347
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TRC8 CRISPR Activation Plasmid (h) | sc-403689-ACT | 20 µg | $397.00 |
Human RNF139 encodes TRC8, a multi-pass endoplasmic reticulum membrane protein that functions as a RING-type E3 ubiquitin ligase and modulates protein quality control and turnover. TRC8 integrates sterol-sensing and ubiquitin-dependent degradation processes, influencing ER-associated degradation and broader proteostasis pathways. Through regulation of substrate ubiquitination, RNF139 can impact cellular programs linked to growth control, stress responses, and membrane protein homeostasis. Altered RNF139/TRC8 activity has been studied in the context of genomic rearrangements and dysregulated ubiquitin signaling observed in cancer biology and related cell-state transitions.
TRC8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RNF139 expression without altering the underlying DNA sequence.
TRC8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RNF139 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 RNF139 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TRC8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RNF139 locus and enabling the study of TRC8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TRC8 pathway restoration in tumor cells with silenced or reduced RNF139 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.