LRRC59 CRISPR Activation Plasmid (h): sc-404821-ACT

LRRC59 (leucine rich repeat containing 59) encodes an endoplasmic reticulum and nuclear envelope–associated membrane protein implicated in protein targeting and membrane-associated trafficking at the cytoplasmic face of the ER. LRRC59 has been linked to ribosome recruitment and co-translational processes that support secretory and membrane protein biogenesis, connecting it to ER homeostasis and proteostasis networks. Through its localization at the ER–nuclear envelope interface, LRRC59 is also studied in the context of nucleo-cytoplasmic organization and stress-responsive remodeling of the endomembrane system. Dysregulated ER proteostasis and altered trafficking programs are recurrent features in cancer and neurodegeneration research, making LRRC59 a useful node for dissecting pathway-level changes in these disease-relevant processes.

LRRC59 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LRRC59 expression without altering the underlying DNA sequence.

LRRC59 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LRRC59 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 LRRC59 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LRRC59 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LRRC59 locus and enabling the study of LRRC59-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LRRC59 pathway restoration in tumor cells with silenced or reduced LRRC59 expression.

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