CAF-1 p60 CRISPR Activation Plasmid (h): sc-405091-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CAF-1 p60 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- CAF-1 p60 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 CAF-1 p60 CRISPR Activation Plasmid (h) and CAF-1 p60 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the CHAF1B 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: CAF-1 p60 Antibody (SS 53): sc-56646
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CAF-1 p60 CRISPR Activation Plasmid (h) | sc-405091-ACT | 20 µg | $397.00 |
CHAF1B encodes the p60 subunit of chromatin assembly factor 1 (CAF-1), a histone chaperone that deposits H3–H4 onto newly synthesized DNA during replication-coupled nucleosome assembly. Through coordination with PCNA at replication forks and during nucleotide excision repair, CAF-1 supports chromatin re-establishment, epigenetic inheritance, and genome stability following DNA synthesis and damage. Disruption of CHAF1B-regulated chromatin maturation can alter replication stress responses, DNA repair fidelity, and transcriptional programs linked to proliferation and differentiation. Aberrant CAF-1 activity has been associated with dysregulated cell cycle control and genome maintenance pathways relevant to cancer biology and other disorders characterized by chromatin instability.
CAF-1 p60 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHAF1B expression without altering the underlying DNA sequence.
CAF-1 p60 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHAF1B 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 CHAF1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CAF-1 p60 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHAF1B locus and enabling the study of CAF-1 p60-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CAF-1 p60 pathway restoration in tumor cells with silenced or reduced CHAF1B expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.