Thyroglobulin CRISPR Activation Plasmid (h): sc-401985-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Thyroglobulin CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Thyroglobulin 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 Thyroglobulin CRISPR Activation Plasmid (h) and Thyroglobulin CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the TG 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: Thyroglobulin Antibody (D-9): sc-365997
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Thyroglobulin CRISPR Activation Plasmid (h) | sc-401985-ACT | 20 µg | $397.00 | |||
Thyroglobulin CRISPR Activation Plasmid (h2) | sc-401985-ACT-2 | 20 µg | $397.00 |
TG encodes thyroglobulin, a large secreted glycoprotein produced by thyroid follicular cells that serves as the primary scaffold for thyroid hormone biosynthesis. Within the follicular lumen, iodination and coupling of tyrosyl residues on thyroglobulin support production of T3 and T4 and integrate with iodine transport, thyroperoxidase activity, and TSH-driven thyroid differentiation programs. TG expression and processing are linked to endoplasmic reticulum protein folding/quality control and vesicular trafficking that govern thyroglobulin secretion and colloid homeostasis. Dysregulation of TG is associated with inherited defects in thyroid hormone synthesis and is relevant to studies of thyroid dysfunction, autoantigen biology, and follicular cell identity.
Thyroglobulin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TG expression without altering the underlying DNA sequence.
Thyroglobulin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TG 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 TG transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Thyroglobulin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TG locus and enabling the study of Thyroglobulin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Thyroglobulin pathway restoration in tumor cells with silenced or reduced TG expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.