KIF1B CRISPR Activation Plasmid (h): sc-403727-ACT

KIF1B encodes a kinesin-3 family microtubule motor that supports ATP-dependent anterograde transport of membranous organelles and cargoes, contributing to axonal trafficking, mitochondrial distribution, and vesicle movement. Through regulation of microtubule-based transport, KIF1B helps maintain neuronal polarity and cellular homeostasis, processes that intersect with pathways controlling mitochondrial dynamics and intracellular signaling. Altered KIF1B expression or function has been linked to neurodevelopmental and neurodegenerative phenotypes and has also been studied in the context of tumor biology, reflecting its role in cell survival and stress responses. These properties make KIF1B a relevant target for dissecting transport-dependent mechanisms underlying nervous system function and disease-associated cellular states.

KIF1B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KIF1B expression without altering the underlying DNA sequence.

KIF1B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KIF1B 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 KIF1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KIF1B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KIF1B locus and enabling the study of KIF1B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KIF1B pathway restoration in tumor cells with silenced or reduced KIF1B expression.

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