Myosin Vb CRISPR Activation Plasmid (h): sc-403324-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Myosin Vb CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Myosin Vb 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 Myosin Vb CRISPR Activation Plasmid (h) and Myosin Vb CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the MYO5B transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Myosin Vb CRISPR Activation Plasmid (h) | sc-403324-ACT | 20 µg | $397.00 |
MYO5B encodes the actin-based motor protein myosin Vb, a key regulator of intracellular membrane trafficking that coordinates recycling endosome dynamics and polarized transport. Myosin Vb interacts with Rab GTPases to direct vesicle movement and cargo delivery, supporting apical–basolateral polarity, tight junction maintenance, and receptor recycling in epithelial cells. Disruption of MYO5B-dependent trafficking perturbs brush border organization and membrane protein localization, processes central to intestinal epithelial homeostasis. MYO5B dysfunction is strongly linked to congenital enteropathies such as microvillus inclusion disease and related cholestatic phenotypes, making it relevant for studies of epithelial polarity and endosomal sorting.
Myosin Vb CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MYO5B expression without altering the underlying DNA sequence.
Myosin Vb CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MYO5B 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 MYO5B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Myosin Vb expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MYO5B locus and enabling the study of Myosin Vb-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Myosin Vb pathway restoration in tumor cells with silenced or reduced MYO5B expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.