CRISPR-mediated Stxbp1 gene activation restores Munc18-1 levels in Stxbp1-haploinsufficient mice

CRISPR-ON using dCas9-VPR restored endogenous Stxbp1 expression and normalized Munc18-1 protein to wild-type levels in neonatal Stxbp1+/- mice.

• Why it matters: STXBP1 haploinsufficiency drives severe epileptic encephalopathies, and no curative therapy exists; restoring physiological wild-type expression could offer benefit.
• What they did: In Neuro2A cells, a multiplexed four-gRNA cassette targeting the Stxbp1 promoter increased Stxbp1 mRNA ~six-fold; AAV-PHP.eB delivered this cassette plus Cre to neonatal Stxbp1+/-/dCas9-VPRfl/+ mice.
• The result: Brain Stxbp1 mRNA rose to 1.5-fold of wild-type and Munc18-1 protein returned to wild-type; exploratory seizure measures tended lower but were not statistically tested, and body weight was not rescued.