MBNL2 CRISPR Activation Plasmid (h): sc-401921-ACT

Human MBNL2 (muscleblind-like splicing regulator 2) is an RNA-binding protein that controls alternative pre-mRNA splicing, mRNA localization, and transcript stability by recognizing YGCY-rich motifs in target RNAs. It helps coordinate tissue- and development-specific isoform programs that influence cytoskeletal organization, cell differentiation, and stress-responsive RNA processing. Dysregulation of MBNL family activity is closely linked to aberrant splice-switching observed in myotonic dystrophy and other repeat-expansion disorders, and altered MBNL2-dependent splicing networks have been associated with neuromuscular and neurodevelopmental phenotypes. Modulating MBNL2 levels is therefore useful for dissecting splicing-factor circuitry, RNA metabolism pathways, and isoform-specific mechanisms in human cellular models.

MBNL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MBNL2 expression without altering the underlying DNA sequence.

MBNL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MBNL2 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 MBNL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MBNL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MBNL2 locus and enabling the study of MBNL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MBNL2 pathway restoration in tumor cells with silenced or reduced MBNL2 expression.

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