Expanding CRISPR repertoire using CjCas9 as a smaller editing tool

Area of Science:

• Molecular Biology
• Gene Therapy
• Biotechnology

Background:

• CRISPR-Cas9 technology is advancing rapidly for treating genetic diseases.
• The large size of SpCas9 limits its delivery via Adeno-Associated Virus (AAV).
• Smaller Cas9 orthologues offer potential for improved viral vector packaging.

Discussion:

• This study focuses on CjCas9, a smaller Cas9 variant suitable for AAV delivery.
• The APOC3 locus was targeted for its predicted cardioprotective effects in humans.
• A humanized mouse model was engineered to study in vivo gene targeting due to the absence of the APOC3 locus in native mice.

Key Insights:

• CjCas9 demonstrates high on-target gene editing efficiency.
• Successful in vivo targeting of the APOC3 locus was achieved in the humanized mouse model.
• Investigated gene integration stability, promoter activity, and long-term therapeutic efficacy.

Outlook:

• CjCas9 holds promise for AAV-mediated gene therapy of genetic disorders.
• Further research will elucidate the long-term safety and efficacy of CjCas9 gene editing.
• This approach could pave the way for novel treatments for cardiovascular diseases.