NKLAM CRISPR Activation Plasmid (h): sc-409031-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- NKLAM CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- NKLAM 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 NKLAM CRISPR Activation Plasmid (h) and NKLAM CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the RNF19B transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
NKLAM CRISPR Activation Plasmid (h) | sc-409031-ACT | 20 µg | $397.00 |
Human RNF19B encodes NKLAM, a RING-type E3 ubiquitin ligase that promotes substrate ubiquitination and proteasome-dependent regulation of immune and stress-responsive proteins. NKLAM activity has been linked to innate immune effector functions, including modulation of antimicrobial responses, cytokine signaling, and cellular activation programs through ubiquitin-mediated control of signaling components. By shaping protein turnover and signal amplitude, RNF19B participates in pathways that influence inflammation, host–pathogen interactions, and cell fate decisions. Altered regulation of ubiquitination networks involving NKLAM is therefore relevant to studies of immune dysregulation and inflammation-associated disease mechanisms.
NKLAM CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RNF19B expression without altering the underlying DNA sequence.
NKLAM CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RNF19B 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 RNF19B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NKLAM expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RNF19B locus and enabling the study of NKLAM-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NKLAM pathway restoration in tumor cells with silenced or reduced RNF19B expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.