CRISPR/Cas9 mediated somatic gene therapy for insertional mutations: the vibrator mouse model

Area of Science:

• Neuroscience
• Genetics
• Biotechnology

Background:

• Somatic gene therapy for central nervous system (CNS) disorders faces challenges in gene delivery efficiency and therapeutic efficacy.
• CRISPR/Cas9 technology has shown potential in correcting genetic mutations, but its application for insertional mutations in the brain is less explored.

Purpose of the Study:

• To investigate the efficacy of CRISPR/Cas9-mediated gene therapy in removing a recessive insertional mutation (vibrator, vb) in the mouse brain.
• To evaluate the therapeutic potential of this approach for treating neurodegenerative conditions caused by insertional mutations.

Main Methods:

• Utilized adeno-associated virus (AAV) vectors to deliver Cas9 plasmid and guide RNAs targeting the vibrator mutation in the mouse brain.
• Assessed the impact of the CRISPR/Cas9 treatment on phosphatidylinositol transfer protein, α (PITPα) expression, neurodegeneration, tremor, and survival rates in vb mice.

Main Results:

• CRISPR/Cas9 treatment effectively removed the endogenous retroviral insertion in intron 4 of the PITPα gene.
• Treated vb mice exhibited delayed neurodegeneration, attenuated tremor symptoms, and successfully bypassed juvenile death, indicating restored PITPα function.
• Demonstrated successful gene editing in the postnatal brain, leading to significant therapeutic benefits.

Conclusions:

• CRISPR/Cas9-mediated gene therapy is a viable strategy for correcting recessive insertional mutations in the postnatal brain.
• This study highlights the potential of in situ gene editing for treating genetic neurological disorders.