LASP-1 CRISPR Activation Plasmid (h): sc-404630-ACT

LASP1 encodes LIM and SH3 domain protein 1 (LASP-1), a cytoskeletal adaptor enriched at focal adhesions, lamellipodia, and membrane-associated actin networks where it coordinates protein–protein interactions that regulate cell shape and motility. Through coupling actin-binding partners to adhesion and signaling complexes, LASP-1 influences processes such as migration, invasion, and dynamic remodeling of the extracellular matrix. LASP1 expression and subcellular localization are frequently studied in the context of epithelial–mesenchymal transition, metastatic behavior, and remodeling of adhesion signaling pathways. Dysregulated LASP1 activity has been associated with altered cytoskeletal organization and proliferative signaling in multiple tumor types, supporting its use as a functional node in cancer cell biology research.

LASP-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous LASP1 expression without altering the underlying DNA sequence.

LASP-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the LASP1 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 LASP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LASP-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native LASP1 locus and enabling the study of LASP-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LASP-1 pathway restoration in tumor cells with silenced or reduced LASP1 expression.

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