CD229 CRISPR Activation Plasmid (h): sc-404895-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CD229 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- CD229 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 CD229 CRISPR Activation Plasmid (h) and CD229 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the LY9 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: CD229 Antibody (HLy 9.1.25): sc-58992
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CD229 CRISPR Activation Plasmid (h) | sc-404895-ACT | 20 µg | $397.00 | |||
CD229 CRISPR Activation Plasmid (h2) | sc-404895-ACT-2 | 20 µg | $397.00 |
LY9 encodes CD229, a SLAM family immunoreceptor expressed predominantly on lymphocytes where it modulates cell–cell interactions and signaling thresholds during immune activation. CD229 participates in immunoregulatory pathways linked to T and B cell receptor signaling, influencing cytokine responses, proliferation, and differentiation through adaptor-dependent signaling cascades. By shaping lymphocyte communication within germinal center and peripheral immune niches, CD229 contributes to immune homeostasis and tolerance. Dysregulated LY9/CD229 expression or signaling has been associated with altered immune phenotypes and has been investigated in the context of immune-mediated and hematologic disease biology.
CD229 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LY9 expression without altering the underlying DNA sequence.
CD229 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LY9 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 LY9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD229 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LY9 locus and enabling the study of CD229-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD229 pathway restoration in tumor cells with silenced or reduced LY9 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.