LZTR1 CRISPR Activation Plasmid (h): sc-404531-ACT

LZTR1 (leucine zipper-like transcription regulator 1) encodes a BTB-Kelch family adaptor protein that localizes to the Golgi and functions within CUL3-based E3 ubiquitin ligase complexes to regulate protein ubiquitination and turnover. It has been implicated in controlling RAS/MAPK pathway output through modulation of RAS family proteins and related signaling nodes that influence proliferation, differentiation, and cellular homeostasis. Altered LZTR1 activity has been associated with neurodevelopmental and tumor-predisposition phenotypes, including Noonan syndrome and schwannomatosis, making it a useful target for studying ubiquitin-mediated signaling control in human cells. As a pathway-linked regulator, LZTR1 supports mechanistic investigations into proteostasis, membrane-associated signaling hubs, and genotype–phenotype relationships.

LZTR1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LZTR1 expression without altering the underlying DNA sequence.

LZTR1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LZTR1 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 LZTR1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LZTR1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LZTR1 locus and enabling the study of LZTR1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LZTR1 pathway restoration in tumor cells with silenced or reduced LZTR1 expression.

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