BAM32 CRISPR Activation Plasmid (h): sc-404873-ACT

DAPP1 (BAM32) is an adaptor protein enriched in hematopoietic lineages that couples phosphoinositide signaling to downstream effector pathways in immune cells. By binding phosphatidylinositol (3,4)-bisphosphate via its pleckstrin homology domain, BAM32 supports PI3K-dependent signal propagation from receptors such as the B cell receptor and other immunoreceptors, influencing calcium flux, MAPK activity, and cytoskeletal remodeling. These functions position DAPP1 as a regulator of lymphocyte activation, antigen-driven responses, and inflammatory signaling networks. Dysregulated BAM32-associated pathways have been implicated in aberrant immune cell signaling and are frequently explored in studies of immune dysfunction and hematologic disease mechanisms.

BAM32 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DAPP1 expression without altering the underlying DNA sequence.

BAM32 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DAPP1 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 DAPP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BAM32 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DAPP1 locus and enabling the study of BAM32-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BAM32 pathway restoration in tumor cells with silenced or reduced DAPP1 expression.

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