Genome editing in the mouse brain with minimally immunogenic Cas9 RNPs

• DOI: 10.1016/j.ymthe.2023.06.019
• Publication Date: 2023-07-04T07:53:05Z
• Authors: Elizabeth C. Stahl; Jennifer K. Sabo; Min Hyung Kang; Ryan Allen; Elizabeth Applegate; Shin Eui Kim; Yoonjin Kwon; Anmol Seth; Nicholas Lemus; Viviana Salinas-Rios; Katarzyna M. Soczek; Marena Trinidad; Linda T. Vo; Chris Jeans; Anna Wozniak; Timothy Morris; Athen Kimberlin; Thomas Foti; David F. Savage; Jennifer A. Doudna
• Keywords: Gene Editing; 570; viral vectors; brain; 610; microglia; neurons; Brain; host immune response; 3. Good health; endotoxin/LPS; Mice; Ribonucleoproteins; CRISPR-Associated Protein 9; genome editing; Animals; non-viral delivery; Original Article; CRISPR-Cas9; CRISPR-Cas Systems; mouse

Transient delivery of CRISPR-Cas9 ribonucleoproteins (RNPs) into the central nervous system (CNS) for therapeutic genome editing could avoid limitations viral vector-based including cargo capacity, immunogenicity, and cost. Here, we tested ability cell-penetrant Cas9 RNPs to edit mouse striatum when introduced using a convection-enhanced system. These transient showed comparable neurons reduced adaptive immune responses relative one formulation delivered AAV serotype 9. The production ultra-low endotoxin protein manufactured at scale further improved innate immunity. We conclude that injection-based minimally immunogenic CRISPR CNS provides valuable alternative virus-mediated editing.