SLP-76 CRISPR Activation Plasmid (h): sc-401421-ACT

LCP2 encodes SLP-76, a cytosolic adaptor protein essential for coupling immunoreceptor engagement to downstream signaling in hematopoietic cells. Upon T cell receptor or Fc receptor stimulation, SLP-76 nucleates multiprotein complexes with LAT, VAV1, ITK, and PLCγ to coordinate phosphorylation cascades that regulate calcium flux, MAPK activation, cytoskeletal remodeling, and transcriptional programs governing lymphocyte activation and differentiation. Through these pathways, SLP-76 influences integrin-mediated adhesion, immune synapse formation, and effector responses, making it a central node for studying antigen receptor signaling dynamics. Dysregulated proximal signaling involving SLP-76-associated networks is relevant to immune dysfunction and inflammatory phenotypes, supporting its use in mechanistic models of immune cell signaling.

SLP-76 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LCP2 expression without altering the underlying DNA sequence.

SLP-76 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LCP2 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 LCP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SLP-76 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LCP2 locus and enabling the study of SLP-76-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SLP-76 pathway restoration in tumor cells with silenced or reduced LCP2 expression.

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