Integrin αL/ITGAL/CD11a CRISPR Activation Plasmid (h): sc-402246-ACT

ITGAL encodes integrin αL (CD11a), which heterodimerizes with integrin β2 (CD18) to form LFA-1, a principal adhesion receptor on leukocytes. LFA-1 mediates firm adhesion and transmigration by binding ICAM family ligands and coordinates outside-in signaling that remodels the actin cytoskeleton, supports immunological synapse formation, and tunes T cell activation thresholds. ITGAL activity is regulated through integrin inside-out signaling downstream of chemokine receptors and TCR signaling, linking it to leukocyte trafficking, antigen recognition, and inflammatory responses. Dysregulated ITGAL/LFA-1 function has been associated with altered immune cell adhesion and signaling in autoimmune and inflammatory disease contexts as well as tumor–immune interactions, making it a useful node for mechanistic immunology studies.

Integrin αL/ITGAL/CD11a CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ITGAL expression without altering the underlying DNA sequence.

Integrin αL/ITGAL/CD11a CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ITGAL 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 ITGAL transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin αL/ITGAL/CD11a expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ITGAL locus and enabling the study of Integrin αL/ITGAL/CD11a-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin αL/ITGAL/CD11a pathway restoration in tumor cells with silenced or reduced ITGAL expression.

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