MTCL1 CRISPR Activation Plasmid (h): sc-403629-ACT

MTCL1 (microtubule crosslinking factor 1) encodes a large coiled-coil cytoskeletal protein that organizes and stabilizes microtubule networks by crosslinking microtubules and promoting their accumulation around the Golgi apparatus. Through its role in microtubule anchoring and non-centrosomal microtubule array formation, MTCL1 contributes to cell polarity, vesicle trafficking, and directed migration, processes tightly linked to cytoskeleton-dependent signaling and epithelial organization. Dysregulated microtubule architecture and Golgi positioning are frequently associated with altered proliferation and invasive behavior, making MTCL1 regulation relevant to studies of cancer cell motility and tissue morphogenesis. MTCL1 is also used as a mechanistic entry point for investigating how cytoskeletal remodeling influences organelle dynamics and stress-adaptive cellular responses.

MTCL1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MTCL1 expression without altering the underlying DNA sequence.

MTCL1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MTCL1 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 MTCL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MTCL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MTCL1 locus and enabling the study of MTCL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MTCL1 pathway restoration in tumor cells with silenced or reduced MTCL1 expression.

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