Eg5 CRISPR Activation Plasmid (h): sc-402276-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Eg5 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Eg5 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 Eg5 CRISPR Activation Plasmid (h) and Eg5 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the 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: Eg5 Antibody (F-1): sc-374212
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Eg5 CRISPR Activation Plasmid (h) | sc-402276-ACT | 20 µg | $397.00 | |||
Eg5 CRISPR Activation Plasmid (h2) | sc-402276-ACT-2 | 20 µg | $397.00 |
Human Eg5 (KIF11) encodes a kinesin-5 microtubule motor that drives centrosome separation and bipolar spindle assembly during mitosis, ensuring accurate chromosome congression and segregation. Eg5 function is integrated with microtubule dynamics and spindle assembly checkpoint control, coupling ATP-dependent motility to proper progression through the G2/M transition. Dysregulated Eg5 activity perturbs spindle integrity and promotes chromosomal instability and aneuploidy, processes frequently examined in proliferative disease biology. As a central mitotic regulator, Eg5 is widely used to study cell-cycle control, spindle mechanics, and genome stability pathways.
Eg5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous expression without altering the underlying DNA sequence.
Eg5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the 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 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Eg5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native locus and enabling the study of Eg5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Eg5 pathway restoration in tumor cells with silenced or reduced expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.