TTLL2 CRISPR Activation Plasmid (h): sc-413521-ACT

TTLL2 (tubulin tyrosine ligase–like 2) encodes a member of the TTL-like family of enzymes implicated in post-translational modification of tubulin through polyglutamylation, a process that tunes microtubule dynamics and the interaction of microtubules with motor proteins and microtubule-associated proteins. By modulating axonemal and cytoplasmic microtubule function, TTLL2 can influence cilia/flagella-related processes, intracellular transport, and cytoskeletal organization. Dysregulated tubulin polyglutamylation has been linked in the literature to altered ciliary motility and neurodevelopmental and neurodegenerative phenotypes, supporting TTLL2 as a useful target for studying microtubule-associated disease mechanisms. Human TTLL2 activity is therefore relevant to pathways governing microtubule stability, ciliary biology, and cell-type–specific differentiation programs.

TTLL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TTLL2 expression without altering the underlying DNA sequence.

TTLL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TTLL2 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 TTLL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TTLL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TTLL2 locus and enabling the study of TTLL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TTLL2 pathway restoration in tumor cells with silenced or reduced TTLL2 expression.

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