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.
Nkx-2.5 CRISPR Activation Plasmid (h): sc-400276-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nkx-2.5 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Nkx-2.5 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 Nkx-2.5 CRISPR Activation Plasmid (h) and Nkx-2.5 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the NKX2-5 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: Nkx-2.5 Antibody (A-3): sc-376565
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Nkx-2.5 CRISPR Activation Plasmid (h) | sc-400276-ACT | 20 µg | $397.00 | |||
Nkx-2.5 CRISPR Activation Plasmid (h2) | sc-400276-ACT-2 | 20 µg | $397.00 |
NKX2-5 encodes the homeobox transcription factor Nkx-2.5, a core regulator of cardiogenesis that controls cardiac progenitor specification, chamber morphogenesis, and conduction system development. Nkx-2.5 integrates with GATA4, TBX5, and MEF2 networks to modulate chromatin and transcriptional programs governing sarcomere assembly, calcium handling, and cardiomyocyte differentiation. Dysregulated NKX2-5 activity or expression is associated with congenital heart defects and cardiac conduction abnormalities, making it a key node for studying transcriptional regulation of cardiac lineage commitment. In human cell models, NKX2-5 perturbation provides a tractable entry point to dissect developmental gene regulatory circuits and disease-relevant electrophysiology phenotypes.
Nkx-2.5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NKX2-5 expression without altering the underlying DNA sequence.
Nkx-2.5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NKX2-5 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 NKX2-5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nkx-2.5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NKX2-5 locus and enabling the study of Nkx-2.5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nkx-2.5 pathway restoration in tumor cells with silenced or reduced NKX2-5 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.