CD300E CRISPR Activation Plasmid (h): sc-415572-ACT

CD300E encodes an immunoreceptor of the CD300 family predominantly expressed on myeloid lineage cells, where it participates in regulating innate immune activation. Engagement of CD300E influences signaling networks that coordinate inflammatory cytokine production, phagocyte activation, and antigen-presenting cell function, interfacing with ITAM-associated pathways and downstream kinase cascades that shape cellular responses to pathogens and tissue damage. By modulating myeloid activation states, CD300E is relevant to studies of immune dysregulation in chronic inflammation, autoimmunity, and tumor-associated myeloid remodeling. Its expression dynamics and signaling output make it a useful molecular handle for dissecting receptor-mediated control of monocytes, macrophages, and dendritic cells.

CD300E CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD300E expression without altering the underlying DNA sequence.

CD300E CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD300E 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 CD300E transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD300E expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD300E locus and enabling the study of CD300E-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD300E pathway restoration in tumor cells with silenced or reduced CD300E expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.