Integrin αE/ITGAE/CD103 CRISPR Activation Plasmid (m2): sc-421166-ACT-2

Mouse Itgae encodes integrin αE (CD103), an αEβ7 integrin adhesion receptor that binds E-cadherin to promote retention and positioning of tissue-resident lymphocytes, particularly CD8⁺ T cells and regulatory T cells, within epithelial and mucosal compartments. ITGAE functions in integrin-mediated signaling and cytoskeletal remodeling, influencing immunological synapse formation, migration, and local immune surveillance in barrier tissues. Altered CD103 expression is frequently used to delineate tissue-resident immune states and is implicated in mechanisms of chronic inflammation, tumor–immune interactions, and transplant-related immune responses in experimental models. Gene editing of Itgae in mouse supports studies of leukocyte trafficking, epithelial–immune cell crosstalk, and functional dissection of αEβ7-dependent pathways by flow cytometry phenotyping, in vivo homing assays, and tissue residency profiling.

Integrin αE/ITGAE/CD103 CRISPR Activation Plasmid (m2) provides a targeted, non-destructive approach to upregulating endogenous Itgae expression without altering the underlying DNA sequence.

Integrin αE/ITGAE/CD103 CRISPR Activation Plasmid (m2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Itgae 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 Itgae transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Integrin αE/ITGAE/CD103 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Itgae locus and enabling the study of Integrin αE/ITGAE/CD103-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Integrin αE/ITGAE/CD103 pathway restoration in tumor cells with silenced or reduced Itgae expression.

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