Nek9 CRISPR Activation Plasmid (h): sc-404971-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nek9 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Nek9 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 Nek9 CRISPR Activation Plasmid (h) and Nek9 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the NEK9 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: Nek9 Antibody (39-7): sc-100401
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Nek9 CRISPR Activation Plasmid (h) | sc-404971-ACT | 20 µg | $397.00 |
NEK9 encodes the serine/threonine kinase Nek9, a core regulator of centrosome function and mitotic progression. Nek9 coordinates spindle assembly and microtubule dynamics by activating downstream NIMA-related kinases and integrating phosphorylation events that govern centrosome separation, mitotic entry, and cytokinesis. Through its roles in cell-cycle control and genome stability, altered NEK9 activity is relevant to studies of chromosomal instability and proliferation-associated phenotypes observed in cancer biology and developmental disorders. NEK9 also intersects with stress-responsive signaling that can influence checkpoint fidelity and mitotic exit.
Nek9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NEK9 expression without altering the underlying DNA sequence.
Nek9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NEK9 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 NEK9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nek9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NEK9 locus and enabling the study of Nek9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nek9 pathway restoration in tumor cells with silenced or reduced NEK9 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.