Optimizing Gene Knockdown And In Vivo Base Editing With Novel CRISPR Systems

By Adrian Oliver, Kate Zulauf, Mike Lewis, Hui-Chia Yu-Kemp, Allie Crawley, Tyler McCullough, Melissa Porter, and Michael Coyle

Expanding the therapeutic reach of CRISPR depends on identifying editing systems that can access more of the genome while delivering potent, durable results. Researchers evaluated three systems with complementary PAM recognition sequences and screened more than 100 guide RNAs across potential splice sites. Approximately 15 candidates produced high levels of the expected disruption in primary human hepatocytes. Editing rates also generally aligned with reductions in target mRNA and protein. Further optimization using non-natural guide RNA modifications improved potency and stability, with the strongest designs achieving more than 80% serum biomarker knockdown in vivo.

Discover how systematic target selection, screening, and guide engineering supported near-complete knockdown of a therapeutically relevant gene.