LAPTM5 CRISPR Activation Plasmid (h): sc-404229-ACT

LAPTM5 (lysosomal protein transmembrane 5) is a hematopoietic-enriched lysosomal/endosomal membrane protein that regulates intracellular trafficking and signal attenuation in immune cells. It participates in vesicle-mediated transport and lysosome-dependent turnover of signaling components, helping shape receptor proximal pathways such as antigen receptor signaling, cytokine responses, and ubiquitin-dependent degradation. Through its roles in endolysosomal organization and immune signaling homeostasis, LAPTM5 is relevant to studies of leukocyte activation, inflammatory regulation, and dysregulated signaling programs observed across hematologic disease contexts. Its expression and function are also leveraged to interrogate how lysosome-linked trafficking influences cell fate decisions and proteostasis in immune lineages.

LAPTM5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LAPTM5 expression without altering the underlying DNA sequence.

LAPTM5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LAPTM5 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 LAPTM5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LAPTM5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LAPTM5 locus and enabling the study of LAPTM5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LAPTM5 pathway restoration in tumor cells with silenced or reduced LAPTM5 expression.

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