CD19 CRISPR Activation Plasmid (h): sc-400719-ACT

Human CD19 is a B cell–restricted transmembrane glycoprotein that functions as a coreceptor within the B cell receptor (BCR) complex, amplifying signaling thresholds and shaping antigen-dependent activation. Through coupling to kinases and adaptor proteins, CD19 modulates PI3K–AKT, MAPK/ERK, and NF-κB pathway activity, influencing proliferation, survival, and differentiation programs in developing and mature B cells. Its expression dynamics help define B cell lineage identity and regulate humoral immune responses, including germinal center reactions and immunoglobulin production. Dysregulated CD19 expression or signaling has been linked to altered B cell homeostasis and is frequently examined in the context of B cell malignancies and immune dysfunction.

CD19 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CD19 expression without altering the underlying DNA sequence.

CD19 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CD19 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 CD19 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CD19 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CD19 locus and enabling the study of CD19-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CD19 pathway restoration in tumor cells with silenced or reduced CD19 expression.

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