Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
RPA 32 kDa subunit CRISPR Activation Plasmid (h): sc-401249-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RPA 32 kDa subunit CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- RPA 32 kDa subunit 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 RPA 32 kDa subunit CRISPR Activation Plasmid (h) and RPA 32 kDa subunit CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the RPA2 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: RPA 32 kDa subunit Antibody (9H8): sc-56770
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RPA 32 kDa subunit CRISPR Activation Plasmid (h) | sc-401249-ACT | 20 µg | $397.00 | |||
RPA 32 kDa subunit CRISPR Activation Plasmid (h2) | sc-401249-ACT-2 | 20 µg | $397.00 |
RPA2 encodes the 32 kDa subunit of replication protein A (RPA), an essential heterotrimeric single-stranded DNA-binding complex that stabilizes ssDNA intermediates during DNA replication, recombination, and repair. By coordinating checkpoint activation and the recruitment of nucleases, helicases, and repair polymerases, RPA2 supports ATR-dependent replication stress signaling and preserves fork integrity during S phase. RPA2 function is tightly linked to genome maintenance pathways including nucleotide excision repair, homologous recombination, and DNA damage response networks that prevent mutational burden. Dysregulation of RPA complex activity and replication stress tolerance has been associated with genomic instability phenotypes relevant to cancer biology and other disorders of DNA repair.
RPA 32 kDa subunit CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RPA2 expression without altering the underlying DNA sequence.
RPA 32 kDa subunit CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RPA2 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 RPA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous RPA 32 kDa subunit expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RPA2 locus and enabling the study of RPA 32 kDa subunit-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of RPA 32 kDa subunit pathway restoration in tumor cells with silenced or reduced RPA2 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.