ALP CRISPR Activation Plasmid (h): sc-401877-ACT

PDLIM3 encodes ALP (actinin-associated LIM protein), a PDZ-LIM family scaffold that localizes to the Z-disc in striated muscle and couples cytoskeletal organization to signaling. By binding α-actinin and actin-associated complexes, ALP helps coordinate sarcomere assembly, mechanotransduction, and protein quality control programs that maintain myofibrillar integrity. PDLIM3-associated network perturbations have been linked to altered cardiac and skeletal muscle structure and function, supporting investigation in pathways relevant to cardiomyopathy and myopathies. In cell and tissue models, modulation of ALP expression is commonly used to probe stress responses, cytoskeletal remodeling, and muscle differentiation states.

ALP CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PDLIM3 expression without altering the underlying DNA sequence.

ALP CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PDLIM3 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 PDLIM3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ALP expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PDLIM3 locus and enabling the study of ALP-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ALP pathway restoration in tumor cells with silenced or reduced PDLIM3 expression.

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