Modeling MPPH syndrome in vivo using Breasi-CRISPR

Area of Science:

• Neuroscience
• Genetics
• Developmental Biology

Background:

• Genetic testing identifies novel variants in neurodevelopmental disorders.
• Current methods for variant functional analysis lack either speed or physiological accuracy.
• A need exists for rapid, in vivo methods to study neurodevelopmental disorder variants.

Purpose of the Study:

• To evaluate Breasi-CRISPR (Brain Easi-CRISPR) for rapid in vivo modeling of monogenic neurodevelopmental disorders.
• To assess the efficiency of Breasi-CRISPR in studying megalencephaly postaxial polydactyly polymicrogyria hydrocephalus (MPPH) syndrome.

Main Methods:

• Utilized Breasi-CRISPR for efficient genome editing of neural precursor cells in vivo.
• Electroporated CRISPR-Cas9 reagents into developing mouse brains.
• Analyzed neurodevelopmental phenotypes 2 days post-editing.

Main Results:

• Breasi-CRISPR rapidly induced MPPH syndrome phenotypes in vivo.
• Observed increased cyclin D2 protein and neural progenitor proliferation.
• Demonstrated efficient modeling of MPPH syndrome within 2 days.

Conclusions:

• Breasi-CRISPR is an efficient and rapid technique for in vivo modeling of neurodevelopmental disorders.
• This method accelerates the functional analysis of patient-derived variants.
• Breasi-CRISPR offers a powerful new tool for neurodevelopmental disorder research.