APRIL CRISPR Activation Plasmid (h): sc-403296-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- APRIL CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- APRIL 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 APRIL CRISPR Activation Plasmid (h) and APRIL CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the TNFSF12-TNFSF13 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: APRIL Antibody (F-5): sc-374673
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
APRIL CRISPR Activation Plasmid (h) | sc-403296-ACT | 20 µg | $397.00 |
TNFSF12-TNFSF13 encodes APRIL (a proliferation-inducing ligand; TNFSF13), a TNF superfamily cytokine that regulates B-cell maturation, immunoglobulin class switching, and plasma cell survival. APRIL signals primarily through TNFRSF13B (TACI) and TNFRSF17 (BCMA) to activate NF-κB and related survival and differentiation programs, shaping humoral immune homeostasis. Aberrant APRIL activity has been linked to dysregulated B-cell responses and inflammatory microenvironments, with relevance to autoimmunity and B-cell–associated malignancy biology. In tissue and tumor contexts, APRIL can modulate immune cell crosstalk and stromal interactions that influence proliferation and survival phenotypes.
APRIL CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TNFSF12-TNFSF13 expression without altering the underlying DNA sequence.
APRIL CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TNFSF12-TNFSF13 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 TNFSF12-TNFSF13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous APRIL expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TNFSF12-TNFSF13 locus and enabling the study of APRIL-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of APRIL pathway restoration in tumor cells with silenced or reduced TNFSF12-TNFSF13 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.