Lsh CRISPR Activation Plasmid (m): sc-420828-ACT
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Lsh CRISPR Activation Plasmid (m) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Lsh CRISPR Activation Plasmid (m) 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 Lsh CRISPR Activation Plasmid (m) and Lsh CRISPR Activation Plasmid (m2) target distinct regulatory regions upstream of the Hells 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: Lsh Antibody (H-4): sc-46665
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Lsh CRISPR Activation Plasmid (m) | sc-420828-ACT | 20 µg | $397.00 | |||
Lsh CRISPR Activation Plasmid (m2) | sc-420828-ACT-2 | 20 µg | $397.00 |
Mouse Hells encodes lymphoid-specific helicase (Lsh), an SNF2-family ATP-dependent chromatin remodeler that regulates nucleosome positioning, higher-order chromatin structure, and genome-wide DNA methylation patterns. Lsh supports maintenance of epigenetic silencing at repetitive elements and influences transcriptional programs during development, differentiation, and cell-cycle progression through chromatin accessibility control. It is functionally linked to pathways governing heterochromatin formation, replication-coupled chromatin assembly, and repression of transposable elements, thereby contributing to genome stability. Dysregulation of HELLS/Lsh-associated epigenetic remodeling has been implicated in aberrant methylation landscapes and chromosomal instability phenotypes relevant to cancer biology and neurodevelopmental research.
Lsh CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Hells expression without altering the underlying DNA sequence.
Lsh CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Hells 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 Hells transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Lsh expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Hells locus and enabling the study of Lsh-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Lsh pathway restoration in tumor cells with silenced or reduced Hells expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.