ACTBL2 CRISPR Activation Plasmid (h): sc-415610-ACT

ACTBL2 encodes actin beta-like 2, a cytoskeletal actin family protein implicated in the organization of filamentous actin networks that support cell shape, adhesion, and motility. Actin dynamics coordinate key processes such as cytokinesis, vesicle trafficking, and mechanotransduction, integrating with pathways regulated by Rho-family GTPases and actin-binding proteins. Although ACTBL2 is less characterized than canonical ACTB, altered actin remodeling is broadly relevant to phenotypes involving aberrant migration, invasion, and tissue architecture. As a result, modulation of ACTBL2 expression provides a useful handle for probing cytoskeletal contributions to stress responses and signaling-dependent changes in cellular morphology.

ACTBL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ACTBL2 expression without altering the underlying DNA sequence.

ACTBL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ACTBL2 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 ACTBL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ACTBL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ACTBL2 locus and enabling the study of ACTBL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ACTBL2 pathway restoration in tumor cells with silenced or reduced ACTBL2 expression.

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