CELSR3 CRISPR Activation Plasmid (h): sc-405116-ACT

Human CELSR3 (cadherin EGF LAG seven-pass G-type receptor 3) encodes an atypical adhesion GPCR with multiple cadherin repeats that mediates cell–cell recognition and polarity cues during tissue patterning. CELSR3 functions in planar cell polarity signaling and coordinated cytoskeletal organization, contributing to neuronal migration, axon guidance, and synaptic circuit assembly. Through interactions with core PCP components, it influences directional signaling and morphogenetic processes in developing and mature tissues. Dysregulated CELSR3 expression or PCP network imbalance has been associated with neurodevelopmental phenotypes and has been reported in datasets spanning neurologic and oncology contexts, supporting its use as a mechanistic node for pathway-focused studies.

CELSR3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CELSR3 expression without altering the underlying DNA sequence.

CELSR3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CELSR3 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 CELSR3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CELSR3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CELSR3 locus and enabling the study of CELSR3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CELSR3 pathway restoration in tumor cells with silenced or reduced CELSR3 expression.

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