CD45RC CRISPR Activation Plasmid (h): sc-400245-ACT

PTPRC encodes CD45RC, an isoform of the leukocyte common antigen (CD45) receptor-type protein tyrosine phosphatase that regulates antigen receptor signaling thresholds in hematopoietic cells. By dephosphorylating key Src family kinases and modulating downstream MAPK, PI3K/AKT, and JAK/STAT pathway activity, CD45RC influences T cell activation, cytokine responses, and immune homeostasis. Alternative splicing of PTPRC generates CD45 isoforms with distinct extracellular domains that shape lymphocyte differentiation and subset-specific function. Dysregulated CD45 signaling and isoform balance are relevant to studies of autoimmunity, inflammatory disease mechanisms, and hematologic malignancy biology where altered immune signaling contributes to pathogenesis.

CD45 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTPRC expression without altering the underlying DNA sequence.

CD45 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTPRC 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 PTPRC transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD45 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTPRC locus and enabling the study of CD45-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD45 pathway restoration in tumor cells with silenced or reduced PTPRC expression.

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