Muskelin CRISPR Activation Plasmid (h): sc-409182-ACT

Human MKLN1 encodes muskelin, a multidomain scaffold protein implicated in organizing cytoskeletal architecture and coordinating intracellular trafficking. Muskelin has been linked to actin-dependent processes, vesicle transport, and regulated turnover of protein complexes, supporting spatial control of signaling and receptor dynamics. Through these roles, MKLN1 can influence pathways governing cell adhesion, migration, and proteostasis, which are frequently remodeled in cancer biology and neurobiological contexts. Dysregulated expression or perturbation of muskelin-associated networks has been explored for impacts on cellular morphology, stress responses, and signaling homeostasis in disease-relevant models.

Muskelin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MKLN1 expression without altering the underlying DNA sequence.

Muskelin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MKLN1 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 MKLN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Muskelin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MKLN1 locus and enabling the study of Muskelin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Muskelin pathway restoration in tumor cells with silenced or reduced MKLN1 expression.

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