Integrin αM/CD11b CRISPR Activation Plasmid (h): sc-400563-ACT

ITGAM encodes integrin αM (CD11b), which heterodimerizes with ITGB2 (CD18) to form the Mac-1/CR3 receptor expressed predominantly on myeloid cells. This adhesion and pattern-recognition receptor regulates leukocyte rolling, firm adhesion, and transmigration, and modulates phagocytosis, degranulation, and oxidative burst through outside-in signaling pathways involving Src family kinases, Syk, PI3K-AKT, and MAPK. Integrin αM/CD11b also recognizes complement-opsonized targets (iC3b) and contributes to immune complex handling and inflammatory cytokine programs. Genetic and expression changes in ITGAM are associated with dysregulated innate immune activation and have been studied in contexts including autoimmunity, chronic inflammation, and tumor-associated myeloid cell biology.

Integrin αM/CD11b CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITGAM expression without altering the underlying DNA sequence.

Integrin αM/CD11b CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITGAM 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 ITGAM transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin αM/CD11b expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITGAM locus and enabling the study of Integrin αM/CD11b-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin αM/CD11b pathway restoration in tumor cells with silenced or reduced ITGAM expression.

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