MACF1 CRISPR Activation Plasmid (h): sc-402923-ACT

MACF1 (microtubule actin crosslinking factor 1) is a large cytoskeletal spectraplakin that coordinates microtubule–actin interactions to control cell polarity, intracellular trafficking, and directed migration. By organizing microtubule plus-end dynamics at the cortex and linking cytoskeletal networks, MACF1 contributes to processes such as neurite outgrowth, epithelial organization, and wound-associated motility. Functional studies place MACF1 in pathways that integrate Wnt/β-catenin signaling with cytoskeletal remodeling and adhesion dynamics, influencing tissue morphogenesis and mechanotransduction. Dysregulation or genetic variation in MACF1 has been associated with neurodevelopmental and neuropsychiatric phenotypes and has been investigated in contexts where altered cytoskeletal organization supports invasive cellular behavior.

MACF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MACF1 expression without altering the underlying DNA sequence.

MACF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MACF1 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 MACF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MACF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MACF1 locus and enabling the study of MACF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MACF1 pathway restoration in tumor cells with silenced or reduced MACF1 expression.

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