Factors Impacting Efficacy of AAV-Mediated CRISPR-Based Genome Editing for Treatment of Choroidal Neovascularization

Sook Hyun Chung, Iris Natalie Mollhoff, Uyen Nguyen, Amy Nguyen, Natalie Stucka, Eric Tieu, Suman Manna, Ratheesh Kumar Meleppat, Pengfei Zhang, Emerald Lovece Nguyen, Jared Fong, Robert Zawadzki, Glenn Yiu

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


Frequent injections of anti-vascular endothelial growth factor (anti-VEGF) agents are a clinical burden for patients with neovascular age-related macular degeneration (AMD). Genomic disruption of VEGF-A using adeno-associated viral (AAV) delivery of clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 has the potential to permanently suppress aberrant angiogenesis, but the factors that determine the optimal efficacy are unknown. Here, we investigate two widely used Cas9 endonucleases, SpCas9 and SaCas9, and evaluate the relative contribution of AAV-delivery efficiency and genome-editing rates in vivo to determine the mechanisms that drive successful CRISPR-based suppression of VEGF-A, using a mouse model of laser-induced choroidal neovascularization (CNV). We found that SpCas9 demonstrated higher genome-editing rates, greater VEGF reduction, and more effective CNV suppression than SaCas9, despite similar AAV transduction efficiency between a dual-vector approach for SpCas9 and single-vector system for SaCas9 to deliver the Cas9 orthologs and single guide RNAs (gRNAs). Our results suggest that successful VEGF knockdown using AAV-mediated CRISPR systems may be determined more by the efficiency of genome editing rather than viral transduction and that SpCas9 may be more effective than SaCas9 as a potential therapeutic strategy for CRISPR-based treatment of CNV in neovascular AMD.

Original languageEnglish (US)
Pages (from-to)409-417
Number of pages9
JournalMolecular Therapy - Methods and Clinical Development
StatePublished - Jun 12 2020


  • age-related macular degeneration
  • AMD
  • angiogenesis
  • Cas9
  • CNV
  • genome editing
  • macular degeneration
  • VEGF
  • VEGFa

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics


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