tricellulin CRISPR Activation Plasmid (h): sc-414734-ACT

MARVELD2 encodes tricellulin, a tetraspanning membrane protein enriched at tricellular tight junctions where three epithelial cells meet. Tricellulin contributes to epithelial and endothelial barrier integrity by organizing junctional complexes and regulating paracellular flux, acting in concert with claudins, occludin, and scaffold proteins such as ZO-1. Through its role in tight junction assembly and polarity, MARVELD2 influences processes including epithelial differentiation, mechanotransduction, and tissue homeostasis. Dysregulation or genetic variation in MARVELD2 has been linked to barrier-related pathophysiology, including syndromic and nonsyndromic forms of hearing loss, making it relevant for studies of sensory epithelia and junctional disease mechanisms.

tricellulin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MARVELD2 expression without altering the underlying DNA sequence.

tricellulin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MARVELD2 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 MARVELD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous tricellulin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MARVELD2 locus and enabling the study of tricellulin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of tricellulin pathway restoration in tumor cells with silenced or reduced MARVELD2 expression.

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