LRCH4 CRISPR Activation Plasmid (m): sc-433102-ACT

Mouse Lrch4 encodes LRCH4, a leucine-rich repeat and calponin homology domain–containing protein thought to function as a cytoskeletal adaptor that links protein–protein interaction modules to actin-associated dynamics. LRCH family proteins have been implicated in regulating cell shape, adhesion, and motility, processes that influence tissue remodeling and immune cell behavior. In mammalian systems, LRCH4 has been connected to innate immune signaling networks, including pathways that interface with Toll-like receptor–dependent responses and inflammatory transcriptional programs. Altered LRCH4 regulation has therefore been investigated in the context of inflammatory phenotypes and immune dysregulation relevant to disease-model studies.

LRCH4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Lrch4 expression without altering the underlying DNA sequence.

LRCH4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Lrch4 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 Lrch4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LRCH4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Lrch4 locus and enabling the study of LRCH4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LRCH4 pathway restoration in tumor cells with silenced or reduced Lrch4 expression.

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