MBNL1 CRISPR Activation Plasmid (m): sc-425296-ACT

Mouse Mbnl1 encodes muscleblind-like splicing regulator 1 (MBNL1), an RNA-binding protein that recognizes YGCY motifs in pre-mRNAs to control alternative splicing, mRNA localization, stability, and polyadenylation. MBNL1 is a central component of post-transcriptional gene regulation networks that shape tissue-specific isoform expression during development, with prominent roles in muscle and neuronal programs. Disruption of MBNL1 activity alters splicing outcomes across pathways involved in cytoskeletal organization, ion handling, and synaptic function, contributing to broad changes in cell state. MBNL1-dependent spliceopathy is strongly associated with myotonic dystrophy biology, making Mbnl1 a widely used target for modeling RNA toxicity and splicing-factor insufficiency in mammalian systems.

MBNL1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Mbnl1 expression without altering the underlying DNA sequence.

MBNL1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Mbnl1 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 Mbnl1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MBNL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Mbnl1 locus and enabling the study of MBNL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MBNL1 pathway restoration in tumor cells with silenced or reduced Mbnl1 expression.

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