LEPROTL1 CRISPR Activation Plasmid (h): sc-405575-ACT

LEPROTL1 (leptin receptor overlapping transcript-like 1) encodes a small transmembrane protein implicated in regulating receptor trafficking and endosomal sorting, influencing the abundance and signaling competence of cell-surface receptors. By modulating internalization and recycling, LEPROTL1 can shape downstream pathways linked to growth factor responsiveness and cytokine signaling, with broader effects on cellular metabolism and proliferative control. Altered expression of LEPROTL1 has been reported across molecular profiling studies in human disease contexts, supporting its use as a mechanistic node for investigating dysregulated receptor homeostasis. This makes LEPROTL1 relevant for studies of membrane protein turnover, signal attenuation, and pathway rewiring in stress and disease-associated states.

LEPROTL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LEPROTL1 expression without altering the underlying DNA sequence.

LEPROTL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LEPROTL1 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 LEPROTL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LEPROTL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LEPROTL1 locus and enabling the study of LEPROTL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LEPROTL1 pathway restoration in tumor cells with silenced or reduced LEPROTL1 expression.

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