OBSL1 CRISPR Activation Plasmid (h): sc-407284-ACT

OBSL1 (obscurin-like protein 1) encodes a large cytoskeletal adaptor protein that associates with the actin–myosin network and contributes to intracellular structural organization and mechanotransduction. It is implicated in maintaining cell shape, adhesion dynamics, and cytoskeletal remodeling processes that influence migration and tissue integrity. OBSL1 participates in protein–protein interaction networks that coordinate signaling and scaffold assembly, linking cytoskeletal architecture to broader regulatory pathways controlling growth and differentiation. Genetic disruption of OBSL1 is associated with 3M syndrome and related growth phenotypes, making it relevant for investigating cytoskeleton-linked developmental mechanisms and disease-associated cellular stress responses.

OBSL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous OBSL1 expression without altering the underlying DNA sequence.

OBSL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the OBSL1 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 OBSL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous OBSL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native OBSL1 locus and enabling the study of OBSL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of OBSL1 pathway restoration in tumor cells with silenced or reduced OBSL1 expression.

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