Shuttle Peptide Delivers Base Editor RNPs to Rhesus Monkey Airway Epithelial Cells In Vivo.

Area of Science:

• Gene therapy and genetic engineering
• Respiratory medicine
• Molecular biology

Background:

• Cystic fibrosis (CF) gene editing is hindered by airway epithelial barriers.
• Previous S10 peptide facilitated CRISPR-Cas ribonucleoprotein (RNP) delivery.
• Optimization of S10 led to the development of the S315 peptide for improved base editor RNP delivery.

Approach:

• Intratracheal aerosol delivery of Cy5-labeled S315 peptide in rhesus macaques confirmed respiratory tract distribution.
• Co-administration of ABE8e-Cas9 RNP and S315 targeted the CCR5 gene, achieving up to 5.3% editing efficiency in rhesus airway epithelia.
• ABE8e-Cas9 targeting the CFTR R553X mutation was delivered to cultured human airway epithelial cells.

Key Points:

• S315 peptide enables efficient delivery of base editor RNPs to airway epithelia.
• Gene editing demonstrated persistence in mouse airway epithelia for up to 12 months.
• Functional correction of the CFTR R553X mutation was achieved in human airway epithelial cells, restoring anion channel function.

Conclusions:

• The S315 peptide represents a promising delivery vehicle for base editor RNPs in respiratory gene therapy.
• This strategy holds therapeutic potential for functionally correcting CFTR mutations in cystic fibrosis.
• The findings support the advancement of S315-mediated gene editing for treating respiratory genetic disorders.