p70 S6 kinase β CRISPR Activation Plasmid (h): sc-400409-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- p70 S6 kinase β CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- p70 S6 kinase β 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 p70 S6 kinase β CRISPR Activation Plasmid (h) and p70 S6 kinase β CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the RPS6KB2 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: p70 S6 kinase β Antibody (4B11): sc-293269
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
p70 S6 kinase β CRISPR Activation Plasmid (h) | sc-400409-ACT | 20 µg | $397.00 | |||
p70 S6 kinase β CRISPR Activation Plasmid (h2) | sc-400409-ACT-2 | 20 µg | $397.00 |
Human RPS6KB2 encodes p70 S6 kinase β, an AGC-family serine/threonine kinase that functions downstream of PI3K–AKT–mTOR signaling to couple nutrient and growth factor cues to protein synthesis and cell growth. By phosphorylating substrates involved in translation initiation and ribosomal biogenesis, p70 S6 kinase β helps regulate cell size, proliferation, and metabolic adaptation under changing environmental conditions. Dysregulated mTOR/S6K signaling is frequently studied in the context of altered anabolic metabolism and stress responses in cancer and other proliferative disorders, where aberrant translational control and feedback signaling can reshape pathway output. RPS6KB2 activity is also relevant to investigations of insulin signaling, cellular bioenergetics, and cross-talk with MAPK and autophagy-associated processes.
p70 S6 kinase β CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPS6KB2 expression without altering the underlying DNA sequence.
p70 S6 kinase β CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPS6KB2 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 RPS6KB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous p70 S6 kinase β expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPS6KB2 locus and enabling the study of p70 S6 kinase β-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of p70 S6 kinase β pathway restoration in tumor cells with silenced or reduced RPS6KB2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.