Delivery of CRISPR-Cas9 nickase in a single AAV improves contraction of CAG repeats in HD and improves the translational potential of AAV-mediated delivery of Cas9.
Abstract
The best studied Cas9 enzyme, from Streptococcus pyogenes, is thought to be too large to be efficiently packaged into a single adeno-associated virus (AAV) together with a single-guide (sg) RNA cassette, requiring a dual-vector system. Here we have circumvented this limitation and packaged the full CRISPR system into a single AAV by using a single promoter driving the expression of both the Cas9 enzyme and a sgRNA. We compared the ability of the dual and single vector systems to induce contraction of CAG/CTG repeats in the striatum of a Huntington’s disease model. We show that the all-in-one system led to larger contractions of the CAG/CTG repeats. This approach provides a more efficient and less costly approach for in vivo gene editing and improves the translational potential of AAV-mediated delivery of Cas9. One sentence summary Delivery of CRISPR-Cas9 nickase in a single AAV improves contraction of CAG repeats in HD.
Expanded CAG/CTG repeats cause more than 15 different diseases that all remain without a disease-modifying treatment. Because repeat length accounts for most of the variation in disease severity, contracting CAG/CTG repeats presents an attractive therapeutic avenue. Here, we show that the CRISPR-Cas9 nickase targeted t...
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Background/Objectives: CRISPR/Cas-dependent base editing enables the precise correction of genomic sequences by direct modification of DNA bases without creating potentially mutagenic DNA double-strand breaks. Base editing is considered a therapeutic approach for rare monogenic disorders or viral infections. However, m...
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An improved method for efficient generation of multi-KOs using sgRNA DNA strands and the CRISPR/Cas9 system is presented, which offers a simpler, faster, and cost-effective approach for generating multigene knockout clones, with implications for functional genomics, therapeutic development, and synthetic biology.
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